Journal: Frontiers in Oncology

Article Title: REST corepressor 2 contributes to the cell proliferation of endometrial cancer

doi: 10.3389/fonc.2025.1539263

Figure Lengend Snippet: RCOR2 was overexpressed in endometrial cancer. (A, B) RCOR2 mRNA level in adjacent and tumor tissues of endometrial cancer patients was determined with RT-qPCR (n = 174 for each group). Data was shown with mean ± SD. ***p < 0.001 from t test. (C) ROC analysis of the value of RCOR2 mRNA level in diagnosis of endometrial cancer. (D) Representative Western blots of RCOR2 between adjacent and tumor tissues of endometrial cancer patients.

Article Snippet: For RCOR2 overexpression, an adenovirus expressing human RCOR2 (ad-RCOR2) was obtained from Vector Biolabs (Malvern, USA).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Western Blot

Journal: Frontiers in Oncology

Article Title: REST corepressor 2 contributes to the cell proliferation of endometrial cancer

doi: 10.3389/fonc.2025.1539263

Figure Lengend Snippet: RCOR2 mRNA expression and clinicopathological factors in endometrial cancer patients. (A) RCOR2 mRNA expressions between clinical stage of I-II and III-IV from endometrial cancer patients (n = 104 for I-II and n = 70 for III-IV). (D) is the diagnostic value of RCOR2 mRNA expressions for clinical stage. (B) RCOR2 mRNA expressions in endometrial cancer patients without and with lymph node metastasis (n = 109 for non-metastasis and n = 65 for metastasis). (E) is the diagnostic value of RCOR2 mRNA expressions for lymph node metastasis. (C) RCOR2 mRNA expressions between histologic grade of G1-G2 and G3 from endometrial cancer patients (n = 115 for G1-G2 and n = 59 for G3). (F) is the diagnostic value of RCOR2 mRNA expressions for histologic grade. Data was shown with mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 from t test.

Article Snippet: For RCOR2 overexpression, an adenovirus expressing human RCOR2 (ad-RCOR2) was obtained from Vector Biolabs (Malvern, USA).

Techniques: Expressing, Diagnostic Assay

Journal: Frontiers in Oncology

Article Title: REST corepressor 2 contributes to the cell proliferation of endometrial cancer

doi: 10.3389/fonc.2025.1539263

Figure Lengend Snippet: RCOR2 contributed to cell proliferation in endometrial cancer patients. (A-C) mRNA levels of MKI67, CCND1 and PCNA in adjacent and tumor tissues of endometrial cancer patients was determined with RT-qPCR (n = 174 for each group). Data was shown with mean ± SD. ***p < 0.001 from t test. (D-F) Pearson correlation coefficient analysis was employed to analyze the correlations of RCOR2 mRNA with mRNA levels of MKI67, CCND1 and PCNA in endometrial cancer patients.

Article Snippet: For RCOR2 overexpression, an adenovirus expressing human RCOR2 (ad-RCOR2) was obtained from Vector Biolabs (Malvern, USA).

Techniques: Quantitative RT-PCR

Journal: Frontiers in Oncology

Article Title: REST corepressor 2 contributes to the cell proliferation of endometrial cancer

doi: 10.3389/fonc.2025.1539263

Figure Lengend Snippet: RCOR2 regulated cell viability in Ishikawa (ISK) cells. ISK cells were transfected with si-RCOR2 (50 nM) for 48 h. (A) qRT-PCR was used to measure the mRNA levels of RCOR2 and (B) representative Western blots of RCOR2. The expressions were normalized to control (C) . (D) ISK cells were transfected with ad-RCOR2 (MOI50) for 48 h. qRT-PCR was used to measure the mRNA levels of RCOR2 (E) and representative Western blots of RCOR2 (F) . ISK cells were transfected with si-RCOR2 or ad-RCOR2, cell viability was measured by CCK8 at 24, 48, 72 and 96 hours after the transfection (G, H) . Data was shown with mean ± SD. **p < 0.01, ***p < 0.001 from t test.

Article Snippet: For RCOR2 overexpression, an adenovirus expressing human RCOR2 (ad-RCOR2) was obtained from Vector Biolabs (Malvern, USA).

Techniques: Transfection, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: REST corepressor 2 contributes to the cell proliferation of endometrial cancer

doi: 10.3389/fonc.2025.1539263

Figure Lengend Snippet: RCOR2 regulated cell proliferation in ISK cells. ISK cells were transfected with si-RCOR2 (50 nM) or ad-RCOR2 (MOI50), the colony formation assay was conducted 10 days after transfection and the formation of colonies were compared (A, B) . Data was shown with mean ± SD. ***p < 0.001 from t test. ISK cells were transfected with si-RCOR2 or ad-RCOR2 for 48 hours, mRNA levels of MKI67, CCND1 and PCNA were determined with RT-qPCR (C-E) . Data was shown with mean ± SD. *p < 0.05, **p < 0.01. Data was analyzed by Brown-Forsythe ANOVA comparing control and si-RCOR2, control and ad-RCOR2 groups.

Article Snippet: For RCOR2 overexpression, an adenovirus expressing human RCOR2 (ad-RCOR2) was obtained from Vector Biolabs (Malvern, USA).

Techniques: Transfection, Colony Assay, Quantitative RT-PCR, Control

Journal: Frontiers in Oncology

Article Title: REST corepressor 2 contributes to the cell proliferation of endometrial cancer

doi: 10.3389/fonc.2025.1539263

Figure Lengend Snippet: RCOR2 regulated cell proliferation in ISK cells. ISK cells were transfected with si-RCOR2 (50 nM) or ad-RCOR2 (MOI50), western blotting was used to measure the protein expressions of MKI67, CCND1 and PCNA (A) . The expressions were normalized to control (B-D) . Data was shown with mean ± SD. *p < 0.05, **p < 0.01. Data was analyzed by Brown-Forsythe ANOVA comparing control and si-RCOR2, control and ad-RCOR2 groups.

Article Snippet: For RCOR2 overexpression, an adenovirus expressing human RCOR2 (ad-RCOR2) was obtained from Vector Biolabs (Malvern, USA).

Techniques: Transfection, Western Blot, Control

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A ) Mass spectrometry analysis of RCOR2 protein levels in human tissues. Data were retrieved from ProteomicsDB. ( B ) mRNA expression analysis of RCOR2 across various types of human tumors and normal tissues from TCGA. P values were calculated by unequal-variance t test. Data were retrieved from UALCAN. N/A, not applicable; NS, not significant. ( C ) Single-cell RNA-Seq analysis of RCOR2 in tumors. Data were retrieved from TISCH2. ( D ) Immunoblot analysis of RCOR2 and actin proteins in normal mammary gland and MMTV-PyMT mammary tumors from mice. ( E and F ) Representative RCOR2 IHC in human triple-negative breast tumors and adjacent benign tissues ( E ); staining is quantified with H-score ( F ). * P < 0.05 by paired 2-tailed Student’s t test. Scale bars: 50 μm. ( G and H ) Kaplan-Meier survival analysis for patients with breast cancer by log-rank test. Patients were divided by median expression levels of RCOR2 mRNA. Data were retrieved from TCGA. iBAQ, intensity-based absolute quantification.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: Mass Spectrometry, Expressing, RNA Sequencing, Western Blot, Staining, Quantitative Proteomics

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Immunoblot ( A ) and weight ( B ) of mammary tumors in MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl mice. ( C ) Immunoblot analysis of RCOR2 protein in parental and RCOR2-KO1 or -KO2 MC38 cells. ( D and E ) Growth of parental and RCOR2-KO1 or -KO2 MC38 tumors in C57BL/6J ( D ) and NSG ( E ) mice. ( F ) Flow cytometry analysis of CD8 + T cells (CD45 + CD3e + CD8 + ), CD4 + T cells (CD45 + CD3e + CD4 + ), B cells (CD45 + B220 + ), regulatory T cells (CD45 + CD3e + CD4 + FOXP3 + ), myeloid-derived suppressor cells (MDSCs; CD45 + CD11b + Gr-1 + ), macrophages (CD45 + CD11b + F4/80 + ), and dendritic cells (CD45 + CD11c + F4/80 – ) in parental and RCOR2-KO1 MC38 tumors ( n = 5). ( G and H ) CD8 and CD4 IHC in MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl tumors ( G ); the percentage of T cells is quantified ( H ) ( n = 4). Scale bars: 25 μm. ( I ) Growth of parental and RCOR2-KO1 MC38 tumors in C57BL/6J mice treated with IgG or anti-CD4 and anti-CD8 neutralizing antibodies. ( J ) Growth of parental and RCOR2-KO1 MC38 tumors in C57BL/6J and CD4-KO mice. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( B ), 2-way ANOVA with Tukey’s test ( I and J ) or Dunnett’s test ( D and E ), and 2-tailed Student’s t test ( F and H ). * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: Western Blot, Flow Cytometry, Derivative Assay

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Tumor-free period ( A ) and mammary tumor number ( B ) of MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl mice. ( C ) Immunoblot analysis of RCOR2 protein in non-tumor-initiating cells (Lin – CD90 – ALDH lo ) and tumor-initiating cells (Lin – CD90 – ALDH hi ) isolated from MMTV-PyMT tumors. ( D and E ) Tumorsphere formation assay of MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl tumors. Representative tumorsphere images are shown in D . Tumorsphere numbers are quantified in E ( n = 5). ( F and G ) Flow cytometry analysis ( F ) and quantification ( G ) of tumor-initiation cells in MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl tumors. Representative gating is shown in F . ALDH hi cells are quantified in G . ( H ) Immunoblot analysis of RCOR2 protein in parental, RCOR2-KO1, and RCOR2-rescue MDA-MB-231 cells. ( I and J ) Mammosphere formation assay of parental, RCOR2-KO1, and RCOR2-rescue MDA-MB-231 cells. Representative mammosphere images are shown in I . Mammosphere numbers are quantified in J ( n = 3). ( K ) Limiting dilution assay of parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells in NSG mice. ( L ) Growth of parental and RCOR2-KO1 or -KO2 MDA-MB-231 tumors in NSG mice. ( M and N ) Tumorsphere formation assay in parental and RCOR2-KO1 or -KO2 MDA-MB-231 tumors. Representative tumorsphere images are shown in M . Tumorsphere numbers are quantified in N ( n = 5). ( O and P ) Aldefluor assay (STEMCELL Technologies) in parental and RCOR2-KO1 or -KO2 MDA-MB-231 tumors. Representative flow cytometry gating is shown in O . ALDH hi cells are quantified in P ( n = 5). Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( B , E , G , and J ) or Dunnett’s test ( N and P ), 2-way ANOVA with Dunnett’s test ( L ), log-rank (Mantel-Cox) test ( A ), and χ 2 test ( K ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bars: 100 μm ( D , I , and M ).

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: Western Blot, Isolation, Tube Formation Assay, Flow Cytometry, Limiting Dilution Assay

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Volcano plots of RCOR2 target genes in MDA-MB-231 cells ( n = 2). ( C and D ) Venn diagrams of RCOR2 activated ( C ) and repressed ( D ) gene numbers in MDA-MB-231 cells ( n = 2). ( E and F ) Reactome pathway analysis of RCOR2 target genes in MDA-MB-231 cells ( n = 2). ( G ) RT-qPCR analysis of indicated mRNAs in parental, RCOR2-KO, and RCOR2-rescue MDA-MB-231 cells ( n = 3). ( H ) Flow cytometry analysis of RNF43 protein in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells. ( I ) RT-qPCR analysis of indicated mRNAs in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells treated with 0.1 ng/mL IFN-γ for 24 hours ( n = 3). ( J ) Immunoblot analysis of indicated proteins in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells treated with 0.1 ng/mL IFN-γ for 24 hours. ( K ) Immunoblot analysis of indicated proteins in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO TUBO cells treated with 1 ng/mL IFN-γ for 24 hours. ( L ) Immunoblot analysis of indicated proteins in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO MC38 cells treated with 5 ng/mL IFN-γ for 24 hours. ( M ) Representative immunostaining of I-A/I-E in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO MC38 cells treated with 5 ng/mL IFN-γ for 24 hours. Scale bars: 10 μm. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( G ) or Dunnett’s test ( I ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: Quantitative RT-PCR, Flow Cytometry, Western Blot, Immunostaining

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) RT-qPCR analysis of indicated mRNAs in MDA-MB-231 cells treated with 50 μM GSK-LSD1 ( A ) or 0.2 μM TSA ( B ) for 48 hours ( n = 3). ( C ) RT-qPCR analysis of indicated mRNAs in parental and LSD1-KO MDA-MB-231 cells treated with or without 0.1 ng/mL IFN-γ for 24 hours ( n = 3). ( D ) Immunoblot analysis of indicated proteins in parental and LSD1-KO MDA-MB-231 cells. ( E ) Immunoblot analysis of indicated proteins in parental, HDAC1-KO, and HDAC2-KO MDA-MB-231 cells. ( F ) RT-qPCR analysis of indicated mRNAs in parental, HDAC1-KO, and HDAC2-KO MDA-MB-231 cells treated with or without 0.1 ng/mL IFN-γ for 24 hours ( n = 3). ( G ) RT-qPCR analysis of indicated mRNAs in MDA-MB-231 cells overexpressing empty vector (EV) or RCOR2 treated with DMSO or 50 μM GSK-LSD1 for 48 hours ( n = 3). OE, overexpression. ( H ) RT-qPCR analysis of indicated mRNAs in MDA-MB-231 cells overexpressing EV or RCOR2 treated with DMSO or 0.2 μM TSA for 24 hours and in combination with 0.1 ng/mL IFN-γ for another 24 hours ( n = 3). ( I and J ) Genome browser snapshots of HA, HDAC1, and LSD1 binding peaks, highlighted in gold-yellow, on RNF43 ( I ) and CIITA ( J ) in control, RCOR2-OE, and RCOR2-KO MDA-MB-231 cells ( n = 2). ( K and L ) ChIP-qPCR assay showing relative H3K4me2 ( K ) and H4K16Ac ( L ) occupancy on RNF43 and CIITA in parental and RCOR2-KO MDA-MB-231 cells. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( G and H ) or Dunnett’s test ( C and F ), 2-way ANOVA with Tukey’s test ( K and L ), and 2-tailed Student’s t test ( A and B ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: Quantitative RT-PCR, Western Blot, Plasmid Preparation, Over Expression, Binding Assay, Control, ChIP-qPCR

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Growth of parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO TUBO ( A ) or MC38 ( B ) tumors in BALB/c or C57BL/6J mice. ( C – E ) CD4 and CD8 IHC in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO MC38 tumors ( C ); the percentage of T cells is quantified ( D and E ; n = 5). Scale bars: 25 μm. ( F ) Scheme of MHC-II KO using CRISPR/Cas9. ( G ) Genotyping of MHC-II KO in parental, RCOR2-KO1, MHCII-KO, and RCOR2/MHCII–DKO MC38 cells. ( H ) Representative immunostaining of I-A/I-E in parental, RCOR2-KO1, MHCII-KO, and RCOR2/MHCII–DKO MC38 cells treated with 5 ng/mL IFN-γ for 24 hours. Scale bars: 10 μm. ( I ) Growth of parental, RCOR2-KO1, MHCII-KO, and RCOR2/MHCII–DKO MC38 tumors in C57BL/6J mice. ( J – L ) CD4 and CD8 IHC in parental, RCOR2-KO1, and RCOR2/MHCII–DKO MC38 tumors ( J ); the percentage of T cells is quantified ( K and L ; n = 5). Scale bars: 25 μm. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( D , E , K , and L ) and 2-way ANOVA with Tukey’s test ( A , B , and I ). ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: CRISPR, Immunostaining

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) CD4 + T cell killing assay in coculture with parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 cells ( n = 3). Representative images are shown in A . Cancer cell death is quantified in B . Scale bars: 50 μm. ( C ) RT-qPCR analysis of indicated mRNAs in CD4 + T cells after coculture with parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 cells ( n = 3). ( D and E ) Flow cytometry analysis of GzmB-expressing CD4 + T cells in parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 tumors ( n = 5). Representative gating is shown in D . The percentage of GzmB-expressing CD4 + T cells is quantified in E . ( F and G ) Flow cytometry analysis of GzmB-expressing CD8 + T cells in parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 tumors ( n = 5). Representative gating is shown in F . The percentage of GzmB-expressing CD8 + T cells is quantified in G . Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( B , C , E , and G ). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: Quantitative RT-PCR, Flow Cytometry, Expressing

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A ) Immunoblot of indicated proteins in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells. ( B ) Immunoblot of indicated proteins in parental, RCOR2-KO1, and RCOR2-rescue MDA-MB-231 cells. ( C ) Immunoblot of indicated proteins in parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells. ( D ) Flow cytometry analysis of RNF43 protein in parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells. ( E ) TOPFlash assay in HEK293T cells transfected with EV or RCOR2 and treated with Wnt3a for 48 hours ( n = 3). ( F ) Mammosphere formation assay of parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells. Representative mammosphere images are shown (left), and mammosphere numbers are quantified (right; n = 3). ( G ) Flow cytometry analysis (left) and quantification (right) of ALDH hi cells in parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells ( n = 3). ( H ) Limiting dilution assay of parental, RCOR2-KO1, and RCOR2/RNF43–DKO1 MDA-MB-231 cells in NSG mice. ( I ) Immunoblot of indicated proteins in parental and RCOR2-KO1 MDA-MB-231 cells treated with DMSO or 1 μM CHIR99021 for 48 hours. ( J ) Mammosphere formation assay of parental and RCOR2-KO1 MDA-MB-231 cells treated with DMSO or 1 μM CHIR99021. Representative mammosphere images are shown (left), and mammosphere numbers are quantified (right; n = 3). ( K ) Flow cytometry analysis (left) and quantification (right) of ALDH hi cells in parental and RCOR2-KO1 MDA-MB-231 cells treated with DMSO or 1 μM CHIR99021 for 48 hours ( n = 3). Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( F , G , J , and K ), 2-way ANOVA with Tukey’s test ( E ), and χ 2 test ( H ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bars: 100 μm.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: Western Blot, Flow Cytometry, TOPFlash assay, Transfection, Tube Formation Assay, Limiting Dilution Assay

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A – J ) Analysis of indicated mRNAs in melanoma from complete responders ( n = 5), partial responders ( n = 10), and non-responders ( n = 13) to anti–PD-1 therapy. RNA-Seq data were retrieved from GSE78220 . ( K ) Spearman’s correlation analysis between RCOR2 and CIITA /MHC-II heavy chain gene mRNAs in 1,156 human cancer cell lines. Data were retrieved from the Cancer Cell Line Encyclopedia (CCLE). ( L ) Spearman’s correlation analysis between RCOR2 and CIITA /MHC-II heavy chain gene mRNAs in 1,210 human tumors. Data were retrieved from the International Cancer Genome Consortium/TCGA Pan-Cancer Analysis of Whole Genomes Consortium at cBioPortal. ( M and N ) Growth of parental and RCOR2-KO1 TUBO ( M ) and MC38 ( N ) tumors in BALB/c and C57BL/6J mice, respectively, to which control IgG or anti–PD-1 antibody was administered. ( O ) A proposed mechanistic model of RCOR2-dependent tumor cell plasticity and immune evasion. Data represent mean ± SEM. P values were determined by 2-way ANOVA with Tukey’s test ( M and N ) and 2-tailed Student’s t test ( A ). * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: The tissues were incubated with a peroxidase block, followed by staining with primary antibody: RCOR2 (1:50; homemade), CD8α (1:400; Cell Signaling Technology, catalog 98941), or CD4 (1:100; Cell Signaling Technology, catalog 25229).

Techniques: RNA Sequencing, Control

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A ) Mass spectrometry analysis of RCOR2 protein levels in human tissues. Data were retrieved from ProteomicsDB. ( B ) mRNA expression analysis of RCOR2 across various types of human tumors and normal tissues from TCGA. P values were calculated by unequal-variance t test. Data were retrieved from UALCAN. N/A, not applicable; NS, not significant. ( C ) Single-cell RNA-Seq analysis of RCOR2 in tumors. Data were retrieved from TISCH2. ( D ) Immunoblot analysis of RCOR2 and actin proteins in normal mammary gland and MMTV-PyMT mammary tumors from mice. ( E and F ) Representative RCOR2 IHC in human triple-negative breast tumors and adjacent benign tissues ( E ); staining is quantified with H-score ( F ). * P < 0.05 by paired 2-tailed Student’s t test. Scale bars: 50 μm. ( G and H ) Kaplan-Meier survival analysis for patients with breast cancer by log-rank test. Patients were divided by median expression levels of RCOR2 mRNA. Data were retrieved from TCGA. iBAQ, intensity-based absolute quantification.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: Mass Spectrometry, Expressing, RNA Sequencing, Western Blot, Staining, Quantitative Proteomics

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Immunoblot ( A ) and weight ( B ) of mammary tumors in MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl mice. ( C ) Immunoblot analysis of RCOR2 protein in parental and RCOR2-KO1 or -KO2 MC38 cells. ( D and E ) Growth of parental and RCOR2-KO1 or -KO2 MC38 tumors in C57BL/6J ( D ) and NSG ( E ) mice. ( F ) Flow cytometry analysis of CD8 + T cells (CD45 + CD3e + CD8 + ), CD4 + T cells (CD45 + CD3e + CD4 + ), B cells (CD45 + B220 + ), regulatory T cells (CD45 + CD3e + CD4 + FOXP3 + ), myeloid-derived suppressor cells (MDSCs; CD45 + CD11b + Gr-1 + ), macrophages (CD45 + CD11b + F4/80 + ), and dendritic cells (CD45 + CD11c + F4/80 – ) in parental and RCOR2-KO1 MC38 tumors ( n = 5). ( G and H ) CD8 and CD4 IHC in MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl tumors ( G ); the percentage of T cells is quantified ( H ) ( n = 4). Scale bars: 25 μm. ( I ) Growth of parental and RCOR2-KO1 MC38 tumors in C57BL/6J mice treated with IgG or anti-CD4 and anti-CD8 neutralizing antibodies. ( J ) Growth of parental and RCOR2-KO1 MC38 tumors in C57BL/6J and CD4-KO mice. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( B ), 2-way ANOVA with Tukey’s test ( I and J ) or Dunnett’s test ( D and E ), and 2-tailed Student’s t test ( F and H ). * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: Western Blot, Flow Cytometry, Derivative Assay

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Tumor-free period ( A ) and mammary tumor number ( B ) of MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl mice. ( C ) Immunoblot analysis of RCOR2 protein in non-tumor-initiating cells (Lin – CD90 – ALDH lo ) and tumor-initiating cells (Lin – CD90 – ALDH hi ) isolated from MMTV-PyMT tumors. ( D and E ) Tumorsphere formation assay of MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl tumors. Representative tumorsphere images are shown in D . Tumorsphere numbers are quantified in E ( n = 5). ( F and G ) Flow cytometry analysis ( F ) and quantification ( G ) of tumor-initiation cells in MMTV-PyMT +/– K14-Cre +/– Rcor2 +/+ , MMTV-PyMT +/– K14-Cre +/– Rcor2 +/fl , and MMTV-PyMT +/– K14-Cre +/– Rcor2 fl/fl tumors. Representative gating is shown in F . ALDH hi cells are quantified in G . ( H ) Immunoblot analysis of RCOR2 protein in parental, RCOR2-KO1, and RCOR2-rescue MDA-MB-231 cells. ( I and J ) Mammosphere formation assay of parental, RCOR2-KO1, and RCOR2-rescue MDA-MB-231 cells. Representative mammosphere images are shown in I . Mammosphere numbers are quantified in J ( n = 3). ( K ) Limiting dilution assay of parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells in NSG mice. ( L ) Growth of parental and RCOR2-KO1 or -KO2 MDA-MB-231 tumors in NSG mice. ( M and N ) Tumorsphere formation assay in parental and RCOR2-KO1 or -KO2 MDA-MB-231 tumors. Representative tumorsphere images are shown in M . Tumorsphere numbers are quantified in N ( n = 5). ( O and P ) Aldefluor assay (STEMCELL Technologies) in parental and RCOR2-KO1 or -KO2 MDA-MB-231 tumors. Representative flow cytometry gating is shown in O . ALDH hi cells are quantified in P ( n = 5). Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( B , E , G , and J ) or Dunnett’s test ( N and P ), 2-way ANOVA with Dunnett’s test ( L ), log-rank (Mantel-Cox) test ( A ), and χ 2 test ( K ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bars: 100 μm ( D , I , and M ).

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: Western Blot, Isolation, Tube Formation Assay, Flow Cytometry, Limiting Dilution Assay

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Volcano plots of RCOR2 target genes in MDA-MB-231 cells ( n = 2). ( C and D ) Venn diagrams of RCOR2 activated ( C ) and repressed ( D ) gene numbers in MDA-MB-231 cells ( n = 2). ( E and F ) Reactome pathway analysis of RCOR2 target genes in MDA-MB-231 cells ( n = 2). ( G ) RT-qPCR analysis of indicated mRNAs in parental, RCOR2-KO, and RCOR2-rescue MDA-MB-231 cells ( n = 3). ( H ) Flow cytometry analysis of RNF43 protein in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells. ( I ) RT-qPCR analysis of indicated mRNAs in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells treated with 0.1 ng/mL IFN-γ for 24 hours ( n = 3). ( J ) Immunoblot analysis of indicated proteins in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells treated with 0.1 ng/mL IFN-γ for 24 hours. ( K ) Immunoblot analysis of indicated proteins in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO TUBO cells treated with 1 ng/mL IFN-γ for 24 hours. ( L ) Immunoblot analysis of indicated proteins in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO MC38 cells treated with 5 ng/mL IFN-γ for 24 hours. ( M ) Representative immunostaining of I-A/I-E in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO MC38 cells treated with 5 ng/mL IFN-γ for 24 hours. Scale bars: 10 μm. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( G ) or Dunnett’s test ( I ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: Quantitative RT-PCR, Flow Cytometry, Western Blot, Immunostaining

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) RT-qPCR analysis of indicated mRNAs in MDA-MB-231 cells treated with 50 μM GSK-LSD1 ( A ) or 0.2 μM TSA ( B ) for 48 hours ( n = 3). ( C ) RT-qPCR analysis of indicated mRNAs in parental and LSD1-KO MDA-MB-231 cells treated with or without 0.1 ng/mL IFN-γ for 24 hours ( n = 3). ( D ) Immunoblot analysis of indicated proteins in parental and LSD1-KO MDA-MB-231 cells. ( E ) Immunoblot analysis of indicated proteins in parental, HDAC1-KO, and HDAC2-KO MDA-MB-231 cells. ( F ) RT-qPCR analysis of indicated mRNAs in parental, HDAC1-KO, and HDAC2-KO MDA-MB-231 cells treated with or without 0.1 ng/mL IFN-γ for 24 hours ( n = 3). ( G ) RT-qPCR analysis of indicated mRNAs in MDA-MB-231 cells overexpressing empty vector (EV) or RCOR2 treated with DMSO or 50 μM GSK-LSD1 for 48 hours ( n = 3). OE, overexpression. ( H ) RT-qPCR analysis of indicated mRNAs in MDA-MB-231 cells overexpressing EV or RCOR2 treated with DMSO or 0.2 μM TSA for 24 hours and in combination with 0.1 ng/mL IFN-γ for another 24 hours ( n = 3). ( I and J ) Genome browser snapshots of HA, HDAC1, and LSD1 binding peaks, highlighted in gold-yellow, on RNF43 ( I ) and CIITA ( J ) in control, RCOR2-OE, and RCOR2-KO MDA-MB-231 cells ( n = 2). ( K and L ) ChIP-qPCR assay showing relative H3K4me2 ( K ) and H4K16Ac ( L ) occupancy on RNF43 and CIITA in parental and RCOR2-KO MDA-MB-231 cells. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( G and H ) or Dunnett’s test ( C and F ), 2-way ANOVA with Tukey’s test ( K and L ), and 2-tailed Student’s t test ( A and B ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: Quantitative RT-PCR, Western Blot, Plasmid Preparation, Over Expression, Binding Assay, Control, ChIP-qPCR

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) Growth of parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO TUBO ( A ) or MC38 ( B ) tumors in BALB/c or C57BL/6J mice. ( C – E ) CD4 and CD8 IHC in parental, RCOR2-KO1, CIITA-KO, and RCOR2/CIITA–DKO MC38 tumors ( C ); the percentage of T cells is quantified ( D and E ; n = 5). Scale bars: 25 μm. ( F ) Scheme of MHC-II KO using CRISPR/Cas9. ( G ) Genotyping of MHC-II KO in parental, RCOR2-KO1, MHCII-KO, and RCOR2/MHCII–DKO MC38 cells. ( H ) Representative immunostaining of I-A/I-E in parental, RCOR2-KO1, MHCII-KO, and RCOR2/MHCII–DKO MC38 cells treated with 5 ng/mL IFN-γ for 24 hours. Scale bars: 10 μm. ( I ) Growth of parental, RCOR2-KO1, MHCII-KO, and RCOR2/MHCII–DKO MC38 tumors in C57BL/6J mice. ( J – L ) CD4 and CD8 IHC in parental, RCOR2-KO1, and RCOR2/MHCII–DKO MC38 tumors ( J ); the percentage of T cells is quantified ( K and L ; n = 5). Scale bars: 25 μm. Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( D , E , K , and L ) and 2-way ANOVA with Tukey’s test ( A , B , and I ). ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: CRISPR, Immunostaining

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A and B ) CD4 + T cell killing assay in coculture with parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 cells ( n = 3). Representative images are shown in A . Cancer cell death is quantified in B . Scale bars: 50 μm. ( C ) RT-qPCR analysis of indicated mRNAs in CD4 + T cells after coculture with parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 cells ( n = 3). ( D and E ) Flow cytometry analysis of GzmB-expressing CD4 + T cells in parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 tumors ( n = 5). Representative gating is shown in D . The percentage of GzmB-expressing CD4 + T cells is quantified in E . ( F and G ) Flow cytometry analysis of GzmB-expressing CD8 + T cells in parental, RCOR2-KO1, RCOR2/CIITA–DKO, and RCOR2/MHCII–DKO MC38 tumors ( n = 5). Representative gating is shown in F . The percentage of GzmB-expressing CD8 + T cells is quantified in G . Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( B , C , E , and G ). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: Quantitative RT-PCR, Flow Cytometry, Expressing

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A ) Immunoblot of indicated proteins in parental and RCOR2-KO1 or -KO2 MDA-MB-231 cells. ( B ) Immunoblot of indicated proteins in parental, RCOR2-KO1, and RCOR2-rescue MDA-MB-231 cells. ( C ) Immunoblot of indicated proteins in parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells. ( D ) Flow cytometry analysis of RNF43 protein in parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells. ( E ) TOPFlash assay in HEK293T cells transfected with EV or RCOR2 and treated with Wnt3a for 48 hours ( n = 3). ( F ) Mammosphere formation assay of parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells. Representative mammosphere images are shown (left), and mammosphere numbers are quantified (right; n = 3). ( G ) Flow cytometry analysis (left) and quantification (right) of ALDH hi cells in parental, RCOR2-KO1, and RCOR2/RNF43–DKO MDA-MB-231 cells ( n = 3). ( H ) Limiting dilution assay of parental, RCOR2-KO1, and RCOR2/RNF43–DKO1 MDA-MB-231 cells in NSG mice. ( I ) Immunoblot of indicated proteins in parental and RCOR2-KO1 MDA-MB-231 cells treated with DMSO or 1 μM CHIR99021 for 48 hours. ( J ) Mammosphere formation assay of parental and RCOR2-KO1 MDA-MB-231 cells treated with DMSO or 1 μM CHIR99021. Representative mammosphere images are shown (left), and mammosphere numbers are quantified (right; n = 3). ( K ) Flow cytometry analysis (left) and quantification (right) of ALDH hi cells in parental and RCOR2-KO1 MDA-MB-231 cells treated with DMSO or 1 μM CHIR99021 for 48 hours ( n = 3). Data represent mean ± SEM. P values were determined by 1-way ANOVA with Tukey’s test ( F , G , J , and K ), 2-way ANOVA with Tukey’s test ( E ), and χ 2 test ( H ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bars: 100 μm.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: Western Blot, Flow Cytometry, TOPFlash assay, Transfection, Tube Formation Assay, Limiting Dilution Assay

Journal: The Journal of Clinical Investigation

Article Title: Tumoral RCOR2 promotes tumor development through dual epigenetic regulation of tumor plasticity and immunogenicity

doi: 10.1172/JCI188801

Figure Lengend Snippet: ( A – J ) Analysis of indicated mRNAs in melanoma from complete responders ( n = 5), partial responders ( n = 10), and non-responders ( n = 13) to anti–PD-1 therapy. RNA-Seq data were retrieved from GSE78220 . ( K ) Spearman’s correlation analysis between RCOR2 and CIITA /MHC-II heavy chain gene mRNAs in 1,156 human cancer cell lines. Data were retrieved from the Cancer Cell Line Encyclopedia (CCLE). ( L ) Spearman’s correlation analysis between RCOR2 and CIITA /MHC-II heavy chain gene mRNAs in 1,210 human tumors. Data were retrieved from the International Cancer Genome Consortium/TCGA Pan-Cancer Analysis of Whole Genomes Consortium at cBioPortal. ( M and N ) Growth of parental and RCOR2-KO1 TUBO ( M ) and MC38 ( N ) tumors in BALB/c and C57BL/6J mice, respectively, to which control IgG or anti–PD-1 antibody was administered. ( O ) A proposed mechanistic model of RCOR2-dependent tumor cell plasticity and immune evasion. Data represent mean ± SEM. P values were determined by 2-way ANOVA with Tukey’s test ( M and N ) and 2-tailed Student’s t test ( A ). * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: NSG, C57BL/6J, BALB/c, Rcor2 fl/fl (B6.129- Rcor2 tm1.1Gman /J), MMTV-PyMT [B6.FVB-Tg(MMTV-PyVT)634Mul/LellJ], and CD4-KO (B6.129S2-Cd4 tm1Mak /J) mice were purchased from The Jackson Laboratory.

Techniques: RNA Sequencing, Control