{"database":"bioimages","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"submitter":["Ming-Xin An"],"journal":["The Journal of Cell Biology"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-JCBD-201701064"],"attach_to":["JCB"],"legend":["A, Representative immunohistochemistry staining with BAG3 and HK2 in pancreatic cancer tissues.<br />C, Western blot was performed using tissue samples from age-matched mouse littermates without (control) or with transgenic Bag3 knock-in (KI).  <br />D, MEF cells were isolated from control or Bag3 KI embryo, and HK2 expression was analyzed using Western blot.","A, Pancreatic cancer tissue chip was immunohistochemically stained using BAG3 antibody. Representative immunohistochemical images were provided<br />D, BxPC3 were infected with gRNA guided BAG3 using CRISPR/Cas9 system, Western blot was performed using the indicated antibodies. <br />H, SW1990 cells were infected with gRNA guided BAG3 using CRISPR/Cas9 system, BAG3 expression was confirmed using Western blot.<br />J, Control or BAG3 knockdown BxPC3 cells were injected intracutaneously into nude mice. Tumors were excised after mice sacrifice on day 28 (n=10 mice/group). Tumors were photographed (upper panel) and weights were measured (lower panel). *, P<0.01.","C, Control or BAG3 knockdown BxPC3 cells were infected with shRNAs against IMP1 (shIMP1) or HSP70 (shHSP70) containing lentivirus, HK2 expression was analyzed using Western blot.<br />I, Control or BAG3 knockdown BxPC3 cells were infected with lentivirus containing scramble or shRNA against Roquin (shRoquin). Roquin and HK2 protein levels were analyzed using Western blot.<br />J, Biotinylated wild-type (WT) or CDE mutation (Mut) HK2 3’UTR mRNA was used to pulldown lysates from BxPC3 cells, and the pulldown materials were analyzed by western blotting analysis.","A, HK2 expression was analyzed using lysates from control or BAG3 knockdown BxPC3 or SW1990 cells. <br />G, BxPC3 or SW1990 cells were infected with lentivirus containing empty or BAG3 construct, HK2 protein was analyzed using Western blot.","D, HEK293 cells were cotransfected with BAG3 and IMP3 constructs. Interaction of BAG3 with IMP3 was analyzed using co-IP and subsequent Western blot.<br />E, co-IP was performed using lysates from BxPC3 or SW1990 cells. <br />F, Direct interaction of endogenous BAG3 with IMP3 was analyzed using DuoLink PLA in BxPC3 or SW1990 cells. <br />J, BxPC3 cells transduced with lentivirus containing empty or BAG3 construct were infected with lentivirus containing scramble or shRNA against IMP3 (shIMP3), Western blot was analyzed using the indicated antibodies.","E, BxPC3 cells were infected with lentivirus containing scramble or shAgo2, and Western blot was performed using the indicated antibodies.","A, BxPC3 or SW1990 cells were infected with lentivirus containing empty or BAG3 construct, BAG3 expression was confirmed using Western blot.","C, BxPC3 cells were infected with lentivirus containing shRNAs against IMP1 (shIMP1) or HSP70 (shHSP70), Western blot was performed using the indicated antibodies.<br />E, Biotinylated RNA segment of HK2 mRNA was used to pulldown lysates from BxPC3 cells, and the pulldown materials were analyzed by western blotting analysis using the indicated antibodies.<br />H-I, BxPC3 cells were infected with lentivirus containing Flag-tagged wild-type (WT) or BAG3 construct with deletion of potential RNA-binding domain. HK2 expression was investigated using  Western blot(H). Interaction of BAG3 and HSP70 or IMP1 was investigated using co-IP (I)."],"repository":["bioimages"],"figure_sub":["Figure 1 - A,D,H,J","Figure 4 - A,G","Figure 5 - C,E,H,I","Figure 8 - A,C,D","Image 642545 (Figure 3 - A)","Image 642548 (Figure 6 - C,I,J)","Figure 3 - A","Image 642550 (Figure 8 - A,C,D)","Figure 5","Figure 7 - D,E,F,J","Figure 4","Figure 7","Figure 6","Figure 6 - C,I,J","Figure 8","Image 642549 (Figure 7 - D,E,F,J)","Image 642544 (Figure 1 - A,D,H,J)","Sup Figure 3 - E","Figure 1","Image 642547 (Figure 5 - C,E,H,I)","Image 642551 (Sup Figure 3 - E)","Figure 3","Sup Figure 3","Image 642546 (Figure 4 - A,G)"],"pubmed_authors":["Hua-Qin Wang","Xin-Yu Li","Si Li","Jia-Mei Wang","Xiao-Na Meng","Chao Li","Ming-Xin An","Jia Sun","Han-Bing Yao"],"additional_accession":[]},"is_claimable":false,"name":"BAG3 directly stabilizes Hexokinase 2 mRNA and promotes aerobic glycolysis in pancreatic cancer cells","description":"Aerobic glycolysis, a phenomenon known historically as the Warburg effect, is one of the hallmarks of cancer cells. In this study, we characterized the role of BAG3 in aerobic glycolysis of pancreatic ductal adenocarcinoma (PDAC) and its molecular mechanisms. Our data show that aberrant expression of BAG3 significantly contributes to the reprogramming of glucose metabolism in PDAC cells. Mechanistically, BAG3 increased Hexokinase 2 (HK2) expression, the first key enzyme involved in glycolysis, at the posttranscriptional level. BAG3 interacted with HK2 mRNA, and the degree of BAG3 expression altered recruitment of the RNA-binding proteins Roquin and IMP3 to the HK2 mRNA. BAG3 knockdown destabilized HK2 mRNA via promotion of Roquin recruitment, whereas BAG3 overexpression stabilized HK2 mRNA","dates":{"release":"2017-11-07T11:29:15Z","modification":"2018-11-29T11:29:15Z","creation":"2018-11-29T11:29:15Z"},"accession":"S-JCBD-201701064","cross_references":{"doi":["10.1083/jcb.201701064"]}}