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Identification of p62-interacting proteins by mass spectrometry analysis in HEK293T cells

ABSTRACT: HEK293T cells were ectopically expressing Flag tagged p62 (p62 group) or empty vectors (Control group) for 48h, cell lysates were incubated with anti-Flag affinity gels, and co-immunoprecipitates were subjected to trypsin digestion followed by mass spectrometry analysis. Peptides were separated by the EASY-nLC system (Thermo Fisher) and analyzed by the Q Exactive mass spectrometer (Thermo Fisher). Protein analysis were performed with Thermo Proteome Discoverer 2.1 (Thermo Fisher) and searched against Uniprot Human database. Three samples per group.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: yun yang

LAB HEAD: yang yun

PROVIDER: PXD061794 | Pride | 2025-07-29

REPOSITORIES: Pride

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			Action	DRS
	YS_CHI_194_P62_1.raw	Raw
	YS_CHI_194_P62_2.raw	Raw
	YS_CHI_194_P62_3.raw	Raw
	YS_CHI_194_V_1.raw	Raw
	YS_CHI_194_V_2.raw	Raw

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Publications

TRIM25 ubiquitinates and degrades p62/SQSTM1 to suppress autophagy.

Qiu Xiang X Ren Jin J Yang Yun Y Hu Weikang W Wang Chengcheng C Hu Ronggui R Li Chuanyin C

Acta biochimica et biophysica Sinica 20250620

PMID: 40542578

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Identification of p62-interacting proteins by mass spectrometry analysis in HEK293T cells

Project description:HEK293T cells were ectopically expressing Flag tagged p62 (p62 group) or empty vectors (Control group) for 48h, cell lysates were incubated with anti-Flag affinity gels, and co-immunoprecipitates were subjected to trypsin digestion followed by mass spectrometry analysis. Peptides were separated by the EASY-nLC system (Thermo Fisher) and analyzed by the Q Exactive mass spectrometer (Thermo Fisher). Protein analysis were performed with Thermo Proteome Discoverer 2.1 (Thermo Fisher) and searched against Uniprot Human database. Three samples per group.

2025-07-29 | PXD062511 | Pride

Urinary exosome microRNA signatures as a non-invasive prognostic biomarker for prostate cancer

Project description:We aimed to identify urinary exosomal miRNAs associated with PCa metastasis and develop a non-invasive risk-scoring model for PCa metastasis in this study. MiRNA profiles were examined using the Taqman low-density miRNA array (TLDA). Megaplex reverse transcription reactions and pre-amplification reactions were performed to increase the quantity of cDNA for miRNA expression analysis using the Megaplex PreAmp Primers Human Pool A and TaqMan PreAmp Master Mix (Thermo Fisher Scientific). MiRNA expression was evaluated via the TLDA panel A v2.0 (Thermo Fisher Scientific). Raw data were processed using the QuantStudio Real-Time PCR Software (Thermo Fisher Scientific) to determine a cycle threshold (Ct) value for each miRNA.

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Thermo Fisher Scientific OncoScan expression array data for undifferentiated uterine sarcomas (CNV)

Project description:Thermo Fisher Scientific OncoScan expression array data for undifferentiated uterine sarcoma

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CIRCULATING SERUM MICRO-RNA AS NON-INVASIVE DIAGNOSTIC BIOMARKERS OF ENDOMETRIOSIS

Project description:Total RNA was extracted from 400 ul of serum with the miRNeasy Serum/Plasma advanced Kit (Qiagen) and quantified using the Qubit microRNA assay kit (Thermo Fisher Scientific). cDNA templates were prepared using the TaqMan Advanced miRNA cDNA Synthesis Kit (Thermo Fisher Scientific), starting from 10 ng of RNA. RT-qPCR carried out on a QuantStudio 12K Flex (Applied Biosystems) using the TaqMan OpenArray miRNA panel.

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Microarray analysis of lung SPC/GFP+ EpCAM+ cells

Project description:We extracted RNA of sorted lung SPC/GFP+ EpCAM+ cells and performed microarray analyses. Total RNA was extracted from sorted Ep-CAMhigh/GFPhigh cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. The RNA integrity was examined on an Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA). Biotinylated ss-cDNA were prepared according to the standard Affymetrix protocol from 100 ng total RNA using the GeneChip WT PLUS Reagent Kit User Manual (Affymetrix/Thermo Fisher Scientific). Fragmented and labeled ss-cDNA were hybridized on a GeneChip Clariom S Array (Affymetrix/Thermo Fisher Scientific) (n =3/group). We used microarrays to detail the global gene expression of SPC/GFP+ EpCAM+ cells from the control mice(G29-5_(Clariom_S_Mouse)) and the 3 months smoke mice(G29-6_(Clariom_S_Mouse)), and identified distinct classes of up-regulated genes during this process.

2019-04-26 | GSE130331 | GEO

microarray analysis of lung SPC/GFP+ EpCAM+ cells from different smoke exposure mice

Project description:We extracted RNA of sorted lung SPC/GFP+ EpCAM+ cells and performed microarray analyses. Total RNA was extracted from sorted Ep-CAMhigh/GFPhigh cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. The RNA integrity was examined on an Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA). Biotinylated ss-cDNA were prepared according to the standard Affymetrix protocol from 100 ng total RNA using the GeneChip WT PLUS Reagent Kit User Manual (Affymetrix/Thermo Fisher Scientific). Fragmented and labeled ss-cDNA were hybridized on a GeneChip Clariom S Array (Affymetrix/Thermo Fisher Scientific) (n =3/group). We used microarrays to detail the global gene expression of SPC/GFP+ EpCAM+ cells from the control mice(G37-1_(Clariom_S_Mouse)), 3 week continuous smoke mice (G37-2_(Clariom_S_Mouse)) and 3 weeks intermittent smoke mice(G37-3_(Clariom_S_Mouse)), and identified distinct classes of up-regulated genes during this process.

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2022-02-25 | PXD009428 | Pride

Elucidating the Role of the Desmosome Protein p53 Apoptosis Effector Related to PMP-22 in Growth Hormone Tumors

Project description:An Affymetrix Human Gene 1.1 ST array plate (Thermo Fisher Scientific) was hybridized with 2.3 µg of fragmented and labeled single-stranded complementary DNA (cDNA)

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The regulons of an OmpR-like transcription factor in Pseudomonas syringae pv. Actinidiae

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2022-12-13 | GSE220275 | GEO

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