Journal: EMBO Molecular Medicine

Article Title: Targeting pre-existing club-like cells in prostate cancer potentiates androgen deprivation therapy

doi: 10.1038/s44321-026-00375-y

Figure Lengend Snippet: ( A , B ) Expression per Pten pc−/− LSC med cell subpopulation ( A ) and UMAP projection ( B ) of Pim1 . ( C , D ) Number ( C ) and size ( D ) of organoids generated from sorted Pten pc−/− LSC med cells after 10 days of culture in medium containing DMSO or 1 nM JQ-1 and 1 nM CX-6258, alone or combined, as indicated. Data were normalized to the control (DMSO) condition (biological replicates, n = 5 independent experiments). ** p < 0.01; *** p < 0.001; **** p < 0.0001 (one-way ANOVA and Dunnett post hoc test). Exact p values are reported in Appendix Table . ( E , F ) Enrichment of various epithelial cell signatures ( E ) and LSC med subpopulation signatures ( F ) in HPV-10, PC-3, DU145, LNCaP, and 22Rv1 cell lines (Data Ref: Wang et al, ). ( G ) Enrichment of CRPC-AR, CRPC-WNT, CRPC-NE, and CRPC-SCL human tumoral subtypes signatures (Dataset ) in HPV-10, PC-3, DU145, LNCaP, and 22Rv1 cell lines. ( H ) Human HPV-10 cells were treated for 72 h with 1 µM JQ-1 and 1 µM CX-6258, alone or combined (as indicated), then the number of adherent cells was counted (biological replicates, n = 4 independent experiments). The data were normalized to the control condition (DMSO). ** p < 0.01; *** p < 0.001 (one-way ANOVA and Dunnett post hoc test). Exact p values are reported in Appendix Table . ( I ) Images of HPV-10 cells after 72 h of treatment. ( J ) The viability of HPV-10 cells (adherent + in suspension) was determined by trypan blue staining (biological replicates, n = 4 independent experiments). ns not significant; * p < 0.05 (one-way ANOVA and Dunnett post hoc test). Exact p values are reported in Appendix Table . ( K , L ) Same as ( H , J ) with PC-3 cells (biological replicates, n = 4 independent experiments). ns not significant; * p < 0.05, ** p < 0.01; *** p < 0.001 (one-way ANOVA and Dunnett post hoc test). Exact p values are reported in Appendix Table . ( M ) Tumorsphere-forming capacity of HPV-10 cells in medium containing DMSO or 1 µM JQ-1 and 1 µM CX - 6258, alone or combined as indicated. Data were normalized to the DMSO condition (biological replicates, n = 3 independent experiments). *** p < 0.001 (one-way ANOVA and Dunnett post hoc test). Exact p values are reported in Appendix Table . All error bars in this figure represent SD. .

Article Snippet: HPV-10 , ATCC , CRL-2220 RRID: CVCL_3495.

Techniques: Expressing, Generated, Control, Suspension, Staining

Journal: EMBO Molecular Medicine

Article Title: Targeting pre-existing club-like cells in prostate cancer potentiates androgen deprivation therapy

doi: 10.1038/s44321-026-00375-y

Figure Lengend Snippet: ( A , B ) Dose-response of JQ-1 ( A ) and CX-6258 ( B ) on the number of organoids formed by sorted Pten pc−/− LSC med cells. Data are normalized to the DMSO condition (biological replicates, n = 5 ( A ) and n = 4 ( B ) independent experiments). **** p < 0.0001 versus DMSO (ANOVA analysis and Dunnett’s post hoc test). Exact p values are reported in Appendix Table . ( C ) Expression of FOSL1 and PIM1 in human HPV-10 cells determined by RT-qPCR. The results are normalized to the values obtained in LNCaP cells, represented by the horizontal dotted line (biological replicates, n = 3 independent experiments). * p < 0.05 versus LNCaP cells (unpaired t -test with Welch’s correction), p = 0.0157 ( FOSL1 ), p = 0.0278 ( PIM1 ). ( D ) Dose-response of JQ-1 and CX-6258 on the number of adherent HPV-10 cells. Data were normalized to the DMSO condition (each dot is the average of three biological replicates). **** p < 0.0001 versus DMSO (ordinary two-way ANOVA with Šídák multiple comparisons test). Exact p values are reported in Appendix Table . ( E – H ) Human PC-3 ( E , F ) and HPV-10 ( G , H ) cells were treated with T5224-PROTAC (‘PROTAC’) at 0, 8, and 24 h. The cells were collected at 48 h. The cell number ( E , G ) and cell viability (adherent + in suspension) ( F , H ) were determined by trypan blue staining (biological replicates, n = 4 independent experiments). The data were normalized to DMSO (control condition). * p < 0.05, ** p < 0.01 (ANOVA analysis and Dunn’s post hoc test). ( I ) Human PC-3 cells were treated with siRNA (siScrambled or three different FOSL1 siRNA, as indicated) for 6 h and the cells were collected at 48 h. The expression of FOSL1 was measured by RT-qPCR (biological replicates, n = 2 independent experiments). The data were normalized to siScrambled (control condition). The three siRNA FOSL1 showed similar efficacy. **** p < 0.0001 (ANOVA analysis and Dunnett’s post hoc test). Exact p values are reported in Appendix Table . ( J – L ) Human PC-3 cells were treated with siScrambled or siFOSL1(1) for 6 h and the cells were collected at 72 h. Cell number ( J ) and cell viability (adherent + in suspension) ( K ) were determined by trypan blue staining. The expression of FOSL1 ( L ) was measured by RT-qPCR. The data are normalized to siScrambled (biological replicates, n = 3 independent experiments). ** p < 0.01, *** p < 0.001, **** p < 0.0001 (two-tailed t -test). Exact p values are reported in Appendix Table . All error bars in this figure represent SD.

Article Snippet: HPV-10 , ATCC , CRL-2220 RRID: CVCL_3495.

Techniques: Expressing, Quantitative RT-PCR, Suspension, Staining, Control, Two Tailed Test

Journal: Science Advances

Article Title: CD8 + T cell targeting of tumor antigens presented by HLA-E

doi: 10.1126/sciadv.adm7515

Figure Lengend Snippet: ( A ) Immunoblot of cell lysates of indicated K562-derived cell lines. Expression of endogenous and transfected HLA-E in K562 cells was demonstrated with two HLA-E–specific antibodies MEM-E/02 and 3D12 ( , , ). Note that nontransfected K562 cells express low amounts of HLA-E . V5 epitope–tagged RhPAP was detected with PAP-specific (MAB6240, R&D Systems) or V5-specific mouse monoclonal antibodies. A GAPDH-specific mAb was used as loading control. ( B ) ICS for IFNγ and TNFα of CD8 + T cells from three 68-1 RhCMV/RhPAP-immunized RM after coincubation with the indicated cells in the presence or absence of VL9 peptide. ( C ) Results from five independent experiments showing the average relative frequency (background-subtracted and normalized to K562-E/PAP) of CD69 and IFNγ and/or TNFα-positive CD8 + T cells from 68-1 RhCMV/RhPAP-immunized RM responding to the indicated cell lines in the presence or absence of VL9. Individual results are shown in data file S3. Statistical significance was determined by paired t test (not significant, P > 0.05). ( D ) Immunoblot for HLA-E and FLAG-tagged acid phosphatases. Lysates of stable cell lines of AA7 cells (= K562 cells deleted for HLA-E; fig. S7) transfected with HLA-E alone or together with human PAP, ACP2, or ACPT were electrophoretically separated and probed with the indicated antibodies by immunoblot. ( E ) ICS for IFNγ and TNFα of CD8 + T cells from three 68-1 RhCMV/RhPAP-immunized RM after coincubation with the indicated cell lines. ( F ) Average frequencies (+SEM) of CD69 and IFNγ and/or TNFα-positive CD8 + T cells responding to the indicated cell lines were background subtracted and normalized to AA7-E/PAP. Individual results are shown in data file S3. Statistical significance was determined using paired t test.

Article Snippet: Following blocking with 5% milk in PBS–0.1% Tween-20 (Thermo Fisher Scientific) (PBS-T), membranes were probed with the following antibodies in 5% milk in PBS-T: anti–MHC-E 3D12 (1:1000; Invitrogen 14-9953-82), anti–MHC-E MEM-E/02 (1:5000; Invitrogen MA1-19304), anti-FLAG epitope (1:5000; Sigma-Aldrich #F3165), anti-V5 epitope (1:500; Invitrogen #37-7500), anti–glyceraldehyde phosphate dehydrogenase (GAPDH; 1:5000; Invitrogen #MA5-15738), anti-HA epitope (1:2000; clone HA-7, Sigma-Aldrich H9658), anti-PAP (1:500; MAB6240, R&D Systems) anti–MHC-I heavy chain (1:5000; HC10, Thermo Fisher Scientific), or anti-RhCMV IE2 (15 μg/ml; clone IIA5.2) ( ).

Techniques: Western Blot, Derivative Assay, Expressing, Transfection, Control, Stable Transfection

Journal: Science Advances

Article Title: CD8 + T cell targeting of tumor antigens presented by HLA-E

doi: 10.1126/sciadv.adm7515

Figure Lengend Snippet: ( A ) Immunoblot of cell lysates of the indicated cell lines for HLA-E, PAP, HLA-Ia, and GAPDH. Antibodies used were MEM-E/02 for HLA-E , HC10 for HLA-Ia heavy chains , and MA5-15738 (Invitrogen) for GAPDH. The anti-PAP antibody (MAB6240 R&D Systems) detected a lower molecular weight band in DU145 cells that is likely nonspecific since DU145 cells are known to be PAP negative ( , ). ( B ) ICS for IFNγ and TNFα production of CD8 + T cells from a 68-1 RhCMV/RhPAP-immunized RM after coincubation with the indicated cell lines in the presence or absence of VL9 peptide. In addition, we inhibited potential MHC-II presentation by adding anti-DR and CLIP peptide to all stimulations. ( C ) Average frequencies (+SEM) of CD69 and IFNγ and/or TNFα-positive CD8 + T cells from 68-1 RhCMV/RhPAP-immunized RM after background subtraction. The number of repeat experiments is indicated. Individual results are shown in data file S3. Statistical significance was calculated using paired Mann-Whitney U test (not significant, P > 0.05).

Article Snippet: Following blocking with 5% milk in PBS–0.1% Tween-20 (Thermo Fisher Scientific) (PBS-T), membranes were probed with the following antibodies in 5% milk in PBS-T: anti–MHC-E 3D12 (1:1000; Invitrogen 14-9953-82), anti–MHC-E MEM-E/02 (1:5000; Invitrogen MA1-19304), anti-FLAG epitope (1:5000; Sigma-Aldrich #F3165), anti-V5 epitope (1:500; Invitrogen #37-7500), anti–glyceraldehyde phosphate dehydrogenase (GAPDH; 1:5000; Invitrogen #MA5-15738), anti-HA epitope (1:2000; clone HA-7, Sigma-Aldrich H9658), anti-PAP (1:500; MAB6240, R&D Systems) anti–MHC-I heavy chain (1:5000; HC10, Thermo Fisher Scientific), or anti-RhCMV IE2 (15 μg/ml; clone IIA5.2) ( ).

Techniques: Western Blot, Molecular Weight, MANN-WHITNEY