Journal: iScience

Article Title: Targeting KIF20A blocks lactylation modification to suppress immune escape in hepatocellular carcinoma

doi: 10.1016/j.isci.2026.115372

Figure Lengend Snippet: Glycolysis inhibition-mediated blockade of H3K18la in HCC cells restores the function of co-cultured CD8 + T cells (A) Schematic diagram of the experimental design. (B) Changes in lactate levels in Huh7 cells after treatment with 2-DG or oxamate. (C–E) Representative WB results showing the levels of Pan Kla and H3K18la in Huh7 cells treated with inhibitors. (F) Lactate levels in DKO cells with or without lactate supplementation. (G and H) WB analysis of the protein levels of LDHA, LDHB, Pan Kla, and H3K18la in DKO cells with or without lactate treatment. H3 and β-actin were used as loading controls for lactylation markers and LDH isoforms, respectively. (I) Colony formation assays show the proliferation of HCC cells (control group vs. DKO group) cultured alone or co-cultured with CD8 + T cells. (J) Quantitative analysis of the proliferation of co-cultured CD8 + T cells by FC. (K) FC analysis of the expression levels of GzMB (L) and IFN-γ (M) in co-cultured CD8 + T cells. All data are shown as mean ± SD. (B, D, E, and G) were analyzed using one-way ANOVA, (J) analyzed by two-way ANOVA, and (K) was analyzed by Student’s t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. Experiments were repeated three times.

Article Snippet: APC Anti-Mouse CD8a Antibody , Elabscience , Cat# E-AB-F1104E; RRID: AB_3740201.

Techniques: Inhibition, Cell Culture, Control, Expressing

Journal: iScience

Article Title: Targeting KIF20A blocks lactylation modification to suppress immune escape in hepatocellular carcinoma

doi: 10.1016/j.isci.2026.115372

Figure Lengend Snippet: Knockdown of LDHA/B inhibits HCC growth by activating CD8 + T cell-mediated antitumor immunity (A) WB analysis of LDHA, LDHB, and β-actin (loading control) expression in control and LDHA/B DKO Hepa 1–6 cells. (B) Representative images of tumors dissected from tumor-bearing mice ( n = 6). Tumor weights (C), tumor growth curves (D), Kaplan-Meier survival curve ( n = 6) (E), and lactate contents in tumor tissues (F) of the control and LDHA/B DKO groups ( n = 6). (G) IHC staining of H3K18la and Pan Kla in tumor tissues (scale bars, 100 μm, n = 6). (H–M) FC analysis of tumor-infiltrating immune cells: TILs were isolated from control and LDHA/B-DKO tumors. FC plots (left) and quantitative statistics (right) show: the proportion of CD3 + CD8 + CTLs (H), the proportion of CD44hiCD62Llo effector memory CTLs (I), and the proportions of intratumoral GzMB + (J), and IFN-γ + (K) CTLs with representative scatterplots ( n = 6). Cell aggregates and debris were first excluded, followed by gating on the target cell populations and markers of interest. Representative FC plots and quantitative data are presented. (L and M) Tumor size and growth curves of mice treated with anti-CD8 antibody to deplete CD8 + T cells or isotype control (IgG) ( n = 6). (N) Mouse body weights at the end of the experiment ( n = 6). Data are presented as mean ± SD. (C–F, and H–M) were analyzed using Student’s t test, and (N) was analyzed by two-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, not significant. All experiments were repeated three times. β-actin served as the loading control for WB experiments.

Article Snippet: APC Anti-Mouse CD8a Antibody , Elabscience , Cat# E-AB-F1104E; RRID: AB_3740201.

Techniques: Knockdown, Control, Expressing, Immunohistochemistry, Isolation

Journal: iScience

Article Title: Targeting KIF20A blocks lactylation modification to suppress immune escape in hepatocellular carcinoma

doi: 10.1016/j.isci.2026.115372

Figure Lengend Snippet: KIF20A regulates PD-L1 expression via c-Myc (A and B) Huh7 and HCCLM3 cells were transfected with KIF20A small interfering RNAs (siRNAs) or KIF20A overexpression (OE) pcDNA3.1 plasmids, respectively. The protein levels of KIF20A, c-Myc, and PD-L1 were measured by WB in Huh7 and HCCLM3 cells. (C) ChIP-PCR was performed to assess the binding of c-Myc to the PD-L1 promoter region in Huh7 cells. (D) Huh7 cells were transfected with c-Myc siRNAs, and the knockdown efficiency of c-Myc was verified by RT-PCR. (E and F) Huh7 and HCCLM3 cells were co-transfected with KIF20A OE plasmids and c-Myc siRNAs. The protein levels of KIF20A, PD-L1, and c-Myc were determined by WB. (G) Huh7 cells were co-transfected with pGL3-PD-L1 or pGL3-basic plasmids, KIF20A OE pcDNA3.1 plasmids, c-Myc siRNAs, and pRL-TK plasmids, followed by luciferase activity assay. (H and I) KIF20A promotes HCC cell immune evasion by regulating PD-L1 expression via c-Myc. HCC cells from different groups were co-cultured with CD8 + T cells. Representative FC plots and quantitative analysis of GzMB and IFN-γ expression in CD8 + T cells co-cultured with Huh7 cells. (J and K) Representative images and quantitative analysis of colony formation in Huh7 cells with or without CD8 + T cell co-culture. Data are presented as mean ± SD. Statistical analyses were performed using Student’s t test (D), one-way ANOVA (C, I), or two-way ANOVA (G, K). ∗∗ p < 0.01 and ∗∗∗ p < 0.001. All experiments were repeated three times.

Article Snippet: APC Anti-Mouse CD8a Antibody , Elabscience , Cat# E-AB-F1104E; RRID: AB_3740201.

Techniques: Expressing, Transfection, Over Expression, Binding Assay, Knockdown, Reverse Transcription Polymerase Chain Reaction, Luciferase, Activity Assay, Cell Culture, Co-Culture Assay

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A IOD of ACAT2 expression in CC tissues and adjacent tissues was examined using immunohistochemical staining ( n = 47 biologically independent samples). IOD of DHCR7 B and MSMO1 C expression in CC patients with high ( n = 27 biologically independent samples) or low ( n = 20 biologically independent samples) expression of ACAT2 was examined using immunohistochemical staining. The number of activated CD8 T cells (CD8A + GZMB + ) D or activated NK cells (CD56 + GZMB + ) E infiltrated in the tumor tissues of patients with high ( n = 27 biologically independent samples) and low ACAT2 ( n = 20 biologically independent samples) expression was detected. Data represent mean ± SEM. Statistical analysis was performed using the paired A or unpaired ( B – E ) t-test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: ACAT2 expression in HCeEpiC and CC cell lines was examined using RT-qPCR A and Western blot analysis B ( n = 5 independent experiments). C ACAT2, DHCR7, and MSMO1 expression in CC cells after infection with Scramble-sh, ACAT2-sh #1, and ACAT2-sh #2 was examined using Western blot analysis ( n = 5 independent experiments). D Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in CC cells ( n = 5 independent experiments). The proliferation of CC cells was examined using CCK8 ( E ) and colony formation assays F (n = 5 independent experiments). G CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 T cells for 6 h, respectively, and the death of CC cells was detected ( n = 5 independent experiments). H IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , B ) or two-way ( C - H ) ANOVA, followed by Tukey’s multiple comparisons test ( A – H ).

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Infection, Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A ACAT2 knockdown efficiency in U14 cells was examined using western blot analysis ( n = 10 independent experiments). B Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells (n = 10 animals). C The images and weight of the tumors harvested on day 21 ( n = 10 animals). D Protein expression of ACAT2, MSMO1, DHCR7, and PCNA in transplanted tumors was examined using western blot analysis ( n = 10 animals). E Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in transplanted tumors ( n = 10 animals). The gating strategy for GZMB + NK cells and CD8 + T cells F and quantification G were analyzed using flow cytometry ( n = 10 animals). H Survival of mice over 60 days after subcutaneous inoculation of U14 cells was analyzed using the log-rank test ( n = 20 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , C , E , G ) or two-way ( B , D ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Knockdown, Western Blot, Expressing, Flow Cytometry

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: The proliferation of CC cells was examined using CCK8 A and colony formation assays B ( n = 5 independent experiments). C CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 + T cells, and the death of CC cells was detected ( n = 5 independent experiments). D IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). E TGF-β1 released by CC cells was examined using ELISA ( n = 5 independent experiments). F PD-L1 expression levels in CC cells were observed using immunofluorescence staining ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the two-way ( A – F ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells ( n = 5 animals). B The images and weight of the tumors harvested on day 21 ( n = 5 animals). The gating strategy for GZMB + NK cells and CD8 + T cells C and quantification D were analyzed using flow cytometry ( n = 5 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( B , D ) or two-way A ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Flow Cytometry