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The  BACH1 expression was deleted by CRISPR/Cas9. As control, cells transfected with a non-targeting sgRNA were used. Additionally, cells were transfected for 48hrs with a non-targeting siRNA, or an siRNA targeting CRL2. All samples are in triplicate. Briefly, 20ug of whole proteins prepared from KP cells (control or sgBach1 or siFem1B KP cells, in biological triplicate) was used for mass spectrometry analysis. Briefly, proteins extract was loaded on NuPAGE™ 4–12% Bis–tris acrylamide gels according to the manufacturer’s instructions (Life Technologies). Running of protein was stopped as soon as proteins stacked in a single band. Protein containing bands were stained with Imperial Blue (Pierce), cut from the gel and digested with high sequencing grade trypsin (Promega) before mass spectrometry analysis. Gel pieces were washed and destained using few steps of 100mM NH4HCO3. Destained gel pieces were shrunk with 100 mM ammonium bicarbonate in 50% acetonitrile and dried at RT. Protein spots were then rehydrated using 10mM DTT in 25 mM ammonium bicarbonate pH 8.0 for 45 min at 56°C. This solution was replaced by 55 mM iodoacetamide in 25 mM ammonium bicarbonate pH 8.0 and the gel pieces were incubated for 30 min at room temperature in the dark. They were then washed twice in 25 mM ammonium bicarbonate and finally shrunk by incubation for 5 min with 25 mM ammonium bicarbonate in 50% acetonitrile. The resulting alkylated gel pieces were dried at room temperature. The dried gel pieces were reswollen by incubation in 25 mM ammonium bicarbonate pH 8.0 supplemented with 12.5 ng/µl trypsin (Promega) for 1h at 4°C and then incubated overnight at 37°C. Peptides were harvested by collecting the initial digestion solution and carrying out two extractions; first in 5% formic acid and then in 5% formic acid in 60% acetonitrile. Pooled extracts were dried down in a centrifugal vacuum system. Samples were reconstituted in 0.1% TFA 4% acetonitrile before mass spectrometry using an Orbitrap Fusion Lumos Tribrid Mass Spectrometer (ThermoFisher Scientific, San Jose, CA) online with an Ultimate 3000RSLCnano chromatography system (ThermoFisher Scientific, Sunnyvale, CA). Peptides were separated at 40°C using a two steps linear gradient (4–20% acetonitrile/H2O; 0.1% formic acid for 110 min and 20‐32% acetonitrile/H2O; 0.1% formic acid for 10 min). An EASY‐Spray nanosource was used for peptide ionization (2,200 V, 275°C). MS was conducted using a data-independent acquisition mode (DIA). Full MS scans were acquired in the range of m/z 375–1,500 at a resolution of 120,000 at m/z 200 and the automatic gain control (AGC) was set at 4.0 × 10E5 with a 50 ms maximum injection time. MS2 spectra were acquired in the Orbitrap with a resolution of 30,000, in the mass range of 200-1800 m/z after isolation of parent ion in the quadrupole and fragmentation in the HCD cell under collision Energy of 30%. DIA parent ion range was from 400 to 1000 m/z divided into 40 windows 16 Da wide and from 1000 to 1500 m/z divided into 10 windows 50 Da wide."],"repository":["Pride"],"modification":[""],"quantification_method":[""],"data_protocol":["For protein identification and quantification, relative intensity-based label-free quantification (LFQ) was processed using the DIA-NN 1.8 algorithm (Demichev et al, 2020). Raw files were searched against the Mus Musculus database extracted from UniProt (date 2023-04-03; 55,260 entries) implemented with a contaminant database (Frankenfield et al JPR 2022). The following parameters were used for searches: (i) trypsin allowing cleavage before proline; (ii) one missed cleavage was allowed; (iii) cysteine carbamidomethylation (+57.02146) as a fixed modification and methionine oxidation (+15.99491) and N-terminal acetylation (+42.0106) as variable modifications; (iv) a maximum of 1 variable modification per peptide allowed; and (v) minimum peptide length was 7 amino acids and a maximum of 30 amino acids. The match between runs option was enabled. The precursor false discovery was set to 1%. DIA-NN parameters were set on double-pass mode for Neural Network classifier, Robust LC High precision for quantification strategy and RT-dependent mode for Cross-run normalization. Library was generated using Smart profiling set up. MS1 and MS2 mass accuracy was automatically calculated for precursor charge fixed between 2 and 4.  Main output file from DIA-NN was further filtered at 1% FDR and LFQ intensity was calculated using our DIAgui package at 1% q-value (https://github.com/marseille-proteomique/DIAgui (Gerault et al., 2024)). The statistical analysis was done with Perseus software (version 1.6.15.0), where proteins were pre-filtered to remove contaminants (Tyanova and Cox, 2018). Relative quantification was calculated using fold changes of LFQ intensity between different conditions (Controle, siFem1B, sgBach1). To determine whether a given detected protein was specifically differential, a two-sample t-test was done using permutation-based FDR-controlled employing 250 permutations."],"omics_type":["Proteomics"],"labhead":["Dr Luca Lignitto"],"instrument_platform":[""],"labhead_affiliation":["CRCM, INSERM, CNRS, Institut Paoli-Calmettes, Aix-Marseille Univ. , 27 Boulevard Leï Roure, 13009, Marseille, France"],"submission_type":["PARTIAL"],"species":["Mus Musculus (mouse)"],"publication":["42086045 Ahmed B, Salaun D, Jordan JB, Daulat A, Pastorello C, Brito T, Yang Y, Josselin E, Byrne D, Betzi S, Pelletier A, Vernerey J, Ferreira L, Castellano R, Audebert S, Camoin L, Pane A, Bechara C, Borg JP, Modesti M, Lignitto L. CRL2&lt;sup&gt;FEM1B&lt;/sup&gt; uses heme to recruit BACH1 for degradation and regulate ferroptosis in lung cancer. Mol Cell. 2026 86(10):1993-2011.e10 10.1016/j.molcel.2026.04.008"],"submitter_mail":["stephane.audebert@inserm.fr"],"submitter_affiliation":["Marseille Proteomic, Centre de Recherche en Cancérologie de Marseille, Inserm UMR1068, CNRS UMR7258, Aix Marseille Université U105, Institut Paoli Calmettes, 27 Boulevard Leï Roure CS30059 13273 Marseille Cedex 09 France"],"submitter_country":["France"],"pubmed_abstract":["Lung cancers frequently increase iron demand to sustain growth, which makes them vulnerable to ferroptosis. While Cullin 2-RING ubiquitin ligases (CRL2s) are critical regulators of stress responses and redox balance, their role in ferroptosis mechanisms remains largely unknown. Here, we identify the E3 ligase CRL2<sup>FEM1B</sup> as a key regulator of the ferroptotic response. CRL2<sup>FEM1B</sup> recruits BTB and CNC homolog 1 (BACH1), a transcriptional regulator of ferroptosis, for degradation by recognizing a degron that is directly formed by the redox-sensing molecule heme. By degrading BACH1 in response to heme, CRL2<sup>FEM1B</sup> acts as a switch that dynamically modulates the transcriptional activation of ferroptosis-protective genes, particularly solute carrier family 7 member 11 (SLC7A11). Loss of CRL2<sup>FEM1B</sup> stabilizes BACH1 and suppresses SLC7A11, thereby sensitizing lung tumor cells to ferroptosis inducers in vitro and in preclinical models. Our findings identify CRL2<sup>FEM1B</sup> as a target to increase the efficacy of ferroptosis inducers in lung cancer treatment and define a broader principle whereby endogenous metabolites regulate protein degradation by enabling substrate-E3 ligase interactions."],"pubmed_title":["CRL2&lt;sup&gt;FEM1B&lt;/sup&gt; uses heme to recruit BACH1 for degradation and regulate ferroptosis in lung cancer."],"pubmed_authors":["Ahmed Bashir B, Salaun Daniele D, Jordan Jack B JB, Daulat Avais A, Pastorello Chiara C, Brito Tarcísio T, Yang Yuying Y, Josselin Emmanuelle E, Byrne Deborah D, Betzi Stephane S, Pelletier Amandine A, Vernerey Julien J, Ferreira Lorene L, Castellano Remy R, Audebert Stephane S, Camoin Luc L, Pane Attilio A, Bechara Cherine C, Borg Jean-Paul JP, Modesti Mauro M, Lignitto Luca L"],"additional_accession":[]},"is_claimable":false,"name":"Title Heme acts as a molecular degrader linking CRL2-FEM1B to substrate proteolysis and ferroptosis regulation in lung cancer","description":"In this project, we attempted to explore the differential proteins profile of lung cancer cells in which the BACH1-CRL2 pathway was manipulated. To this end, we performed DIA-based quantitative proteomics in mouse  lung adenocarcinoma KP cells. Control cells and BACH1 null cells were used. Additionally, the expression of CRL2 was silenced using siRNA. A cell line transfected with an siCTRL was used as negative control. 3 biological replicates were used for each sample group.","dates":{"publication":"2026-06-11","submission":"2025-04-15"},"accession":"PXD062953","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:7719","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2499525","NEWT:2853422","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:108458","NEWT:5936","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:542","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","NEWT:550","NEWT:33941","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:575412","NEWT:28112","NEWT:49012","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821"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