anti bap1  (Santa Cruz Biotechnology)

Journal: Biomedicines

Article Title: Echinacoside Alleviates Cisplatin-Induced Acute Kidney Injury by Regulating Bap1 to Inhibit Ferroptosis

doi: 10.3390/biomedicines14092127

Figure Lengend Snippet: ECH potentially interacts with Bap1 and reduces Bap1 expression. ( A ) Heatmap of ferroptosis-associated proteins from 4D-DIA proteomic data. ( B ) Binding affinity scores of ECH with Bap1 and SAT1. ( C ) 3D docking structure and 2D residue interaction diagram of ECH-Bap1 complex. ( D – K ) Molecular dynamics curves of ECH-Bap1 complex, including RMSD, RMSF, SASA, hydrogen bond, trajectory conformation, interaction energy, ligand distance and residue binding free energy. ( L ) Western blot of Bap1 protein in HK-2 cells. ( M ) Western blot analysis of Bap1 protein in kidney tissues.

Article Snippet: After membrane blocking, primary antibodies targeting KIM-1 (A24493, ABclonal, Wuhan, China), NGAL (A2092, ABclonal, Wuhan, China), GPX4 (A27995, ABclonal, Wuhan, China), Slc7a11 (A25302, ABclonal, Wuhan, China), Bap1 (A6533SP, ABclonal, Wuhan, China) and the internal reference GAPDH (A19056, ABclonal, Wuhan, China) were incubated overnight at 4 °C, followed by incubation with corresponding secondary antibodies.

Techniques: Expressing, Binding Assay, Docking Assay, Residue, Western Blot, Analysis

Journal: Biomedicines

Article Title: Echinacoside Alleviates Cisplatin-Induced Acute Kidney Injury by Regulating Bap1 to Inhibit Ferroptosis

doi: 10.3390/biomedicines14092127

Figure Lengend Snippet: Overexpression of Bap1 largely abrogates renoprotective effects of ECH. ( A ) Relative Bap1 mRNA levels in HK-2 cells. ( B ) Western blot and quantitative analysis of Bap1 protein in HK-2 cells. ( C ) Representative PI staining images of HK-2 cells and quantitative analysis of the PI-positive area. ( D ) Western blot analysis of Bap1, Slc7a11 and GPX4 proteins in HK-2 cells. ** p < 0.01 vs. OE-Ctrl group ( A , B ) or cisplatin group ( C , D ); # p < 0.05 vs. Cisplatin + ECH group.

Article Snippet: After membrane blocking, primary antibodies targeting KIM-1 (A24493, ABclonal, Wuhan, China), NGAL (A2092, ABclonal, Wuhan, China), GPX4 (A27995, ABclonal, Wuhan, China), Slc7a11 (A25302, ABclonal, Wuhan, China), Bap1 (A6533SP, ABclonal, Wuhan, China) and the internal reference GAPDH (A19056, ABclonal, Wuhan, China) were incubated overnight at 4 °C, followed by incubation with corresponding secondary antibodies.

Techniques: Over Expression, Western Blot, Analysis, Staining

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: BAP1 inhibits metastatic colonization of UM cells in a DUB activity-dependent manner. A - C BAP1 inhibited growth of UM cells. Mel270 and Omm2.3 cells stably expressing pIRES empty vector, pIRES-WT BAP1 or pIRES-C91S BAP1 were generated by lentivirus. The expression of BAP1 was determined by Western blotting ( A ). Parallel cells in culture were monitored daily by trypan blue exclusion assay ( B ) and colony formation assay in soft agar system ( C ). Data are mean ± SD. ns, not significant; ***, p < 0.001; ****, p < 0.0001, one-way ANOVA with post hoc comparison by Tukey test. D-E BAP1 suppressed the stemness of UM cells. Mel270 and Omm2.3 cells expressing pIRES empty vector, constructs carrying WT BAP1 or its C91S mutant were subjected to ALDH activity assay by flow cytometer ( D ) and serial melanosphere culture ( E ). Percentages of ALDH + cells were from three independent experiments ( D ). Data are mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, one-way ANOVA, post hoc intergroup comparisons. F-I BAP1 hampered metastatic colonization to liver in UM. Mel270 and Omm2.3 cells were stably overexpressed with lentiviral pLV3 vector, pLV3-WT BAP1, or pLV3-C91S BAP1. The expression of BAP1 was examined by Western blotting ( F ). The cells were intrasplenically injected into NOG mice. The BLI signals were monitored weekly for four weeks. The BLI signals on day 28 were analyzed ( G ). The representative photos of livers bearing UM metastatic nodules are shown ( H ). The numbers of metastatic foci on the liver surface were analyzed. Each dot represents one liver. Data are mean ± SD, ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001, one-way ANOVA, post hoc intergroup comparisons. Representative metastatic foci in liver H&E staining are shown ( I ). Scale bar, 500 μm ( top ); 200 μm ( bottom )

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Activity Assay, Stable Transfection, Expressing, Plasmid Preparation, Generated, Western Blot, Trypan Blue Exclusion Assay, Colony Assay, Comparison, Construct, Mutagenesis, Flow Cytometry, Injection, Staining

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: BAP1 physically interacts with VHL. A Whole cell lysates from 293T cells stably expressing Flag-BAP1 constructs were immunoprecipitated (IP) with anti-Flag, the pellets were subjected to mass spectrometric analysis. The BAP1-bound proteins with high scores were listed in the table. B - C Physical interaction between endogenous proteins of BAP1 and VHL were analyzed by Co-IP in Mel270 and Omm2.3 cells. Whole cell lysates of Mel270 and Omm2.3 cells were immunoprecipitated with IgG, anti-VHL ( B ), or anti-BAP1 ( C ), followed by immunoblotting with anti-BAP1 or anti-VHL as indicated. D The colocalization of BAP1 and VHL was explored by IF staining with anti-BAP1 and anti-VHL in Mel270 and Omm2.3 cells. E The in situ interaction between endogenous BAP1 and VHL was detected by Proximity Ligation Assay. F Purified recombinant GST, GST-BAP1 (WT) and GST-BAP1 (C91S) mutant were incubated with His-VHL in vitro. The physical interaction between BAP1 and VHL was then examined by Western blotting analysis. G - H Schematic diagram of pIRES-Flag-BAP1 WT and its deletion mutants is shown ( G ). 293T cells were transfected with indicated constructs, precipitations were then conducted with S-protein agarose beads and subjected to Western blotting with the indicated antibodies ( H ). I Recombinant pGEX-4T1 GST, GST-BAP1 and GST-BAP1 mutants freshly purified from bacteria were incubated with His-VHL prepared from pET-28a in vitro. The interaction between BAP1 and VHL was then examined by GST pull-down. J - K Schematic diagram of Flag-VHL WT and its deletion mutants ( J ). 293T cells were transfected with indicated constructs. Cell lysates were immunoprecipitated with S-protein agarose beads, and then subjected to Western blotting with the indicated antibodies ( K )

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Stable Transfection, Expressing, Construct, Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Staining, In Situ, Proximity Ligation Assay, Purification, Recombinant, Mutagenesis, Incubation, In Vitro, Transfection, Bacteria

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: BAP1 deubiquitinates VHL in UM cells. A - G BAP1 upregulates VHL at protein level. Mel270 and Omm2.3 cells stably transduced with scramble or two different shRNA fragments targeting BAP1 gene were subjected to Western blotting with anti-VHL ( A ), and qRT-PCR ( B ). Data are mean±SD, one-way ANOVA, post hoc intergroup comparisons. Mel270 cells stably expressing scramble or BAP1 shRNAs were treated with control or MG132, and Western blotting was conducted with the indicated antibodies ( C ). UM cells were transfected with pIRES empty vector, pIRES-Flag-WT BAP1 or pIRES-Flag-C91S BAP1, and the endogenous VHL was analyzed with Western blotting ( D ) and qRT-PCR ( E ). Data are mean±SD, one-way ANOVA, post hoc intergroup comparisons. Mel270 cells ectopically expressing pIRES vector or pIRES-WT BAP1 ( F ) were exposed to CHX (100 μg/mL) for different durations, and harvested for Western blotting analysis with antibodies against VHL and actin, respectively ( G ). H - M BAP1 mediates VHL at K48 through its deubiquitination activity. Mel270 cells ectopically expressing pIRES vector, pIRES-WT BAP1 or pIRES-C91S BAP1 combined with pLV3-Flag-BAP1 in the presence of MG132 for 6 hours, the cell lysates were immunoprecipitated with anti-Flag beads, and the polyubiquitylated VHL protein was detected by anti-ubiquitin antibody ( H ). Mel270 cells were transfected with pCMV-HA-VHL and treated with MG132 for 6 h. Cell lysates were immunoprecipitated with anti-HA conjugated beads and the resultant beads were incubated in a cell-free system with proteins of GST, GST-BAP1 or GST-C91S BAP1 purified from bacteria. The polyubiquitylated VHL protein was detected by anti-ubiquitin antibody ( I ). Mel270 cells stably expressing lentiviral scramble or BAP1 shRNAs were transfected with pLV3-Flag-VHL, and treated with MG132 for 6 h before cell lysates were immunoprecipitated with anti-Flag, and the polyubiquitylated VHL protein was detected by anti-ubiquitin antibody ( J ). 293T cells stably expressing lentiviral scramble or BAP1 shRNAs were transfected with pET-28a-His Ub, pCMV-HA VHL, and treated with MG132 for 6 h. Ni-NTA beads were used to pull down His-tagged ubiquitin, and the polyubiquitylated VHL protein was examined ( K ). 293T cells were co-transfected with pLV3-Flag-VHL and constructs carrying HA Ub or its mutants, the in vivo ubiquitination assay was done by immunoprecipitation with anti-Flag, and the polyubiquitylated VHL protein was detected by anti-ubiquitin antibody ( L ). 293T cells were co-transfected with pLV3-Flag-VHL and constructs carrying HA-K48 Ub, HA-K63 Ub in the presence of pIRES-WT BAP1 or pIRES-C91S BAP1, followed by the in vivo ubiquitination assay ( M )

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Stable Transfection, Transduction, shRNA, Western Blot, Quantitative RT-PCR, Expressing, Control, Transfection, Plasmid Preparation, Activity Assay, Immunoprecipitation, Ubiquitin Proteomics, Incubation, Purification, Bacteria, Construct, In Vivo

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: BAP1 inhibits metastatic colonization to liver via VHL. A Luciferase labled-Mel270 and -Omm2.3 cells were transduced with scramble or lentiviral shRNA targeting two different fragments of BAP1. The cells harboring shRNA fragment #2 were ectopically expressed with pIRES-Flag-VHL. The expression of BAP1 and VHL were assessed by Western blotting with antibodies as indicated. B-C BAP1 inhibits the growth of UM cells via VHL. After depletion of BAP1 by shRNA, Mel270-luc and Omm2.3-luc cells were transfected with pIRES-Flag-VHL. The growth of the cells was evaluated by trypan blue exclusion assay ( B ) and colony formation ( C ). Data are mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001, one-way ANOVA, post hoc intergroup comparisons. D-E BAP1 eliminates the UM CSC-like cells via VHL. Mel270-luc and Omm2.3-luc cells in A were subjected to flow cytometer analysis for ALDH + percentage ( D ) and serial culture of tumorsphere ( E ). Data are mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001, one-way ANOVA, post hoc intergroup comparisons. F-H Mel270-luc and Omm2.3-luc cells described in A were intrasplenically injected into NOG mice. The spread of UM cells in vivo was monitored by BLI signals ( F ). The BLI signals on the last day were compared. Data are mean ± SD. *, p < 0.05; ***, p < 0.001, one-way ANOVA, post hoc intergroup comparisons. The nodules on liver surface are shown and the number of nodules on liver surface were analyzed ( G ). Data are mean ± SD. ***, p < 0.001; ****, p < 0.0001, one-way ANOVA, post hoc intergroup comparisons. Metastatic foci in liver sections were analyzed by H&E staining ( H ). Scale bar, 500 μm ( top ); 200 μm ( bottom )

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Luciferase, Transduction, shRNA, Expressing, Western Blot, Transfection, Trypan Blue Exclusion Assay, Flow Cytometry, Injection, In Vivo, Staining

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: BAP1 is phosphorylated at Ser369 by CDK6/4. A Immunoprecipitation of cell lysates from Mel270 or Omm2.3 with IgG or anti-BAP1 antibody was immunoblotted with antibodies against CDK4 and CDK6, respectively. B His-CDK4 or His-CDK6 were expressed by pET-28a plasmid in bacteria. The recombinant GST and GST-BAP1 proteins purified from bacteria expressing pGEX-4T1 constructs were in vitro incubated with the recombinant His-CKD6 ( left ) or His-CDK4 ( right ) protein purified from bacteria expressing pET-28a constructs. The protein interaction between BAP1 and CDK6/4 was analyzed by GST pull-down assay. C-D 293T cells were transfected with pIRES and pIRES-BAP1-Flag. Cell lysates were immunoprecipitated with S-protein agarose beads and subjected to immunoblotting analysis with antibodies against CDK4, CDK6 ( C ) and the antibody specific to pan-phospho-CDK substrates ( D ), respectively. E 293T cells were transfected with pIRES and pIRES-Flag-WT BAP1, and pIRES-Flag-BAP1 SA mutants. Cell lysates were immunoprecipitated with S-protein agarose beads and subjected to immunoblotting with the antibody specific to phospho-CDK substrates. F Mel270 cells transfected with plasmids encoding pLV3, pLV3-Flag-BAP1 WT and pLV3-Flag-BAP1 S369A mutant were treated with LY2835219 (10 µmol/L) for 24 h. Cell lysates were immunoprecipitated with S-protein agarose beads and subjected to immunoblotting with the antibody specific to the phosphorylated BAP1 at S369. G Phosphorylation of BAP1 at Ser369 retards polyubiquitylation of VHL. 293T cells stably transduced with scramble or BAP1-shRNA were transfected with pLV3-HA-VHL and pIRES-BAP1 [WT or its mutants (S369A, S369D)], followed by incubation with MG132 for 6 h. Cell lysates were immunoprecipitated with HA beads and subjected to Western blotting analysis with anti-ubiquitin for the polyubiquitylated VHL protein. H Phosphorylation of BAP1 at Ser369 elevated the expression of VHL. After transduced with scramble or BAP1 shRNA, Mel270-luc and Omm2.3-luc cells were restored with pIRES-BAP1 WT or its mutants (S369A, S369D). Cell lysates were subjected to Western blotting analysis with anti-BAP1 and anti-VHL, respectively. I-J After transduced with lentiviral scramble or shCDK6/4, whole cell lysates ( I ) and exosomes ( J ) from HCT116 cells were then subjected to Western blotting with the specific antibodies against CDK6 and CDK4, respectively. K After exposed to exosomes purified from the CDK6/4-depleted HCT116 cells for 24 h, HCT116 cells were subjected to Western blotting analysis. L-N HCT116 cells were cultured in the presence of TGF-β1 for 24 h, the EMT cells were analyzed ( L ). Data are mean ± SD, *, p < 0.05; ***, p < 0.001, one-way ANOVA, post hoc intergroup comparisons. Whole cell lysates ( M ) and exosomes ( N ) from HCT116 cells were subjected to Western blotting analysis. O MC38 cells were cultured in complete medium, or pH 6.5 complete medium, or 5% O 2 for 48 h, lysates of exosomes fractionated in the culture medium were subjected to Western blotting

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Immunoprecipitation, Plasmid Preparation, Bacteria, Recombinant, Purification, Expressing, Construct, In Vitro, Incubation, Pull Down Assay, Transfection, Western Blot, Mutagenesis, Phospho-proteomics, Stable Transfection, Transduction, shRNA, Ubiquitin Proteomics, Cell Culture

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: CDK6/4 promotes liver colonization in a BAP1-dependent manner in UM. A Mel270-luc and Omm2.3-luc cells were transduced with lentiviral shRNA against CDK4 and CDK6 combined with shRNA against BAP1. The endogenous proteins were analyzed by Western blotting with the indicated antibodies. B-C Mel270-luc and Omm2.3-luc cells stably transduced with shRNA against CDK6/4 and BAP1 were subjected to ALDH activity assay ( B ) and serial melanosphere culture ( C ). Data are mean ± SD. ***, p < 0.001; ****, p < 0.0001, one-way ANOVA, post hoc intergroup comparisons. D-F Mel270 and Omm2.3 cells established in A were intrasplenically inoculated in NOG mice. BLI signals were dynamically monitored ( D , left ) and the curve of BLI signal intensity ( D , right ) were plotted against days after inoculation. Data are mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001, one-way ANOVA, post hoc intergroup comparisons. The nodules on liver surface and the number of nodules on liver surface were analyzed ( E ). Data are mean ± SD. ns, not significant; **, p < 0.01, ****, p < 0.0001, one-way ANOVA, post hoc intergroup comparisons. Metastatic foci in liver sections were analyzed by H&E staining ( F ). Scale bar, 500 μm ( top ); 200 μm ( bottom )

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Transduction, shRNA, Western Blot, Stable Transfection, Activity Assay, Staining

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: BAP1 impedes hepatic colonization of UM cells via VHL. A -C Inhibition of CDK6/4 activity escalates the expression of endogenous VHL protein. Mel270 and Omm2.3 cells were incubated with increasing concentrations of Palbociclib for 24 h. The expression of BAP1 and VHL was analyzed by Western blotting ( A ) and qRT-PCR ( B ), respectively. Mel270 cells were incubated with 100 mg/ml of CHX for durations as indicated in the presence of Palbociclib (10.0 µmol/L, 16 h) or not. Cell lysates were then subjected to Wester blotting analysis with anti-VHL ( C ). D Inhibition of CDK 4/6 decreases the ubiquitination of VHL in Mel270 cells. Mel270 cells transfected with constructs of His-Ub and HA-VHL were exposed to Palbociclib (10.0 µmol/L) for 18 h in presence of MG132. His-tagged ubiquitin was immunoprecipitated with Ni-NTA beads and subjected to immunoblotting analysis with anti-HA. E-F Palbociclib inhibits the growth of UM cells. UM cells and the normal retinal pigment epithelia (RPE) cell line ARPE-19 were treated with increasing concentrations of Palbociclib for 72 h. Cell viability was analyzed by MTS assay. IC 50 values are 15.88 µmol/L for 92.1, 11.67 µmol/L for Mel270, 19.97 µmol/L for Omm2.3, 15.48 µmol/L for Omm1, and 100 µmol/L for ARPE-19, respectively ( E ). After treated with Palbociclib for 24 h, UM cells were harvested and inoculated in double layer soft agar culture for colony formation assay. IC 50 values are 2.59 µmol/L for 92.1, 2.51 µmol/L for Mel270, 3.35 µmol/L for Omm2.3, and 3.71 µmol/L for Omm1, respectively ( F ). G-H Palbociclib eliminates the self-renewal of UM stem-like cells. After incubated with Palbociclib (5.0 µmol/L, 24 h), Mel270 and Omm2.3 cells were subjected to flow cytometer analysis for ALDH + percentage ( G ) and serial culture of tumorsphere ( H ). Data are mean ± SD, **, p < 0.01; ***, p < 0.001; ****, p <0.0001, Student’s t test. I-L Palbociclib suppresses the hepatic colonization of UM cells. Mel270-luc and Omm2.3-luc cells were intrasplenically injected in NOG mice. The mice were orally administered with control or Palbociclib (100 mg/kg/d) for 4 ~ 5 weeks. The spreading of UM cells was monitored by BLI signals. The representative photos of BLI signals in mice on day 35 are shown ( I ). The curves of BLI signals were plotted against days after treatment. Data are mean ± SD. **, p < 0.01; ***, p < 0.001, comparisons were made by the Student’s t test between treatment on the last day ( J ). The photos of livers bearing Mel270-luc or Omm2.3-luc metastatic nodules are shown. The nodules on the liver surface were compared between placebo and Palbociclib treatment. Data are mean ± SD. ***, p < 0.001; ****, p < 0.0001, Student’s t test ( K ). The metastatic foci of UM cells in liver sections were analyzed by H&E staining ( L ). Scale bar, 500 μm ( top ); 200 μm ( bottom )

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Inhibition, Activity Assay, Expressing, Incubation, Western Blot, Quantitative RT-PCR, Ubiquitin Proteomics, Transfection, Construct, Immunoprecipitation, MTS Assay, Colony Assay, Flow Cytometry, Injection, Control, Staining

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: Phosphorylation of BAP1 at Ser369 inhibits hepatic colonization of UM cells. A-B Phosphorylation of BAP1 at Ser369 suppress stemness of UM cells. After transduced with scramble or BAP1 shRNA, Mel270-luc and Omm2.3-luc cells were restored with pIRES-BAP1 WT or its mutants (S369A, S369D), followed by incubation with Palbociclib (10 µmol/L, 24 h) or not. After washed in PBS, these cells were subjected to ALDH activity assay ( A ) and serial melanosphere culture ( B ). Data are mean ± SD. ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, one-way ANOVA, post hoc intergroup comparisons. C-G Phosphorylation of BAP1 at Ser369 inhibits the hepatic colonization of UM cells. Mel270-luc and Omm2.3-luc cells stably depleted BAP1 by shRNA lentivirus were complemented with plasmids encoding pIRES-BAP1 WT or its mutants (S369A, S369D). These cells were intrasplenically injected into NOG mice. Mice were then orally administered with placebo or Palbociclib (100 mg/kg/d) for ~ 4 weeks. The growth of UM cells was monitored by BLI signals. The photos of BLI signals in mice on week 4 are shown ( C ). The curves of BLI signals were plotted against days after treatment ( D ). Data are mean ± SD. ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, one-way ANOVA, post hoc intergroup comparisons were made between the treatment on the last day ( D ). The photos of livers bearing Mel270-luc or Omm2.3-luc metastatic nodules are shown ( E ). The nodules on liver surface were analyzed. Data are mean ± SD, ns, not significant; ***, p < 0.001; ****, p < 0.0001, one-way ANOVA, post hoc intergroup comparisons ( F ). The metastatic foci in H&E staining of liver sections were analyzed ( G ). Scale bar, 500 μm ( top ); 200 μm ( bottom )

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Phospho-proteomics, Transduction, shRNA, Incubation, Activity Assay, Stable Transfection, Injection, Staining

Journal: Molecular Cancer

Article Title: CDK6/4 inactivates BAP1 deubiquitinase destabilizing VHL to promote metastatic colonization in liver

doi: 10.1186/s12943-026-02650-5

Figure Lengend Snippet: The expression of BAP1 and VHL is negatively correlated with metastasis in UM and colorectal carcinoma. A The expression of CDK6, BAP1, S369 p-BAP1 and VHL were examined in paraffin-embed adjacent normal and cancerous tissue sections with corresponding antibodies by IHC staining. The representative photos of adjacent normal tissues of choroid and UM are shown. Scale bar, 100 μm, Olympus IX71. B The expression of CDK6, BAP1, p-BAP1(S369) and VHL in adjacent normal tissues and cancerous tissues were evaluated by semi-quantification by the percentage of positive cells and the intensity as indicated by IHC score. Each dot represents one sample. Data are mean ± SD, **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, Student’s t test. C Pie charts of UM with differentiated expression contents of CDK6, BAP1, S369 p-BAP1 and VHL are shown. N-L, negative-low. D The correlation of BAP1 and VHL in the specimens of 62 UM patients were analyzed. E The expression of CDK6, BAP1, S369 p-BAP1 and VHL were examined in paraffin-embed adjacent normal and cancerous tissue sections of CRC patients with corresponding antibodies by IHC staining. The representative photos of adjacent normal tissues of colon and CRC are shown. Scale bar, 100 μm, Olympus IX71. F The expression of CDK6, BAP1, S369 p-BAP1 and VHL in adjacent normal tissues and cancerous tissues were evaluated by semi-quantification evaluation as indicated by IHC score. Each dot represents one sample. Data are mean ± SD, *, p < 0.05; **, p < 0.01; ****, p < 0.0001, Student’s t test. G The expression of CDK6, BAP1 and VHL in paired tissues of primary CRC and liver metastasis were compared. Data are mean ± SD, *, p < 0.05; **, p < 0.01; ***, p < 0.001, Student’s t test. H Pie charts of CRC with differentiated expression contents of CDK6, BAP1, S369 p-BAP1 and VHL are shown. N-L, negative-low. I The correlation of BAP1 and VHL in the specimens of 97 CRC patients were analyzed. J The correlation of BAP1, S369 p-BAP and VHL with metastasis-free period in the cohort of CRC patients we collected. **, p < 0.01; ****, p < 0.0001, Log-rank test. K The proposed working model of CDK6/4-BAP1-VHL axis and relevant exosome positive loop in metastatic colonization of UM and CRC in liver

Article Snippet: Antibody against BAP1 (cat#A302-243A, RRID: AB_1731059) was from Bethyl (Montgomery, Texas).

Techniques: Expressing, Immunohistochemistry