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Fig. 5 PNCK knockdown enhances the efficacy of immune checkpoint inhibition therapy. (a) The experimental schedule for the SCC7 cell challenge in the C3H mice tumor model is as follows. Intraperitoneal injections of ICIs were administered at a dose of 200 µg per mouse on the 5th, 8th, and 11th days after tumor implantation. The experiment was terminated on the 23rd day, and the tumors were removed for subsequent analysis. If the tumor volume reached 2000 mm³ during the experiment, euthanasia was performed immediately. (b) Overall tumor growth curves in different groups of mice. Mice were intraperitoneally treated with 200 µg anti-PD-L1 or 200 µg anti-CTLA4 on days 5, 8 and 11 after tumor inoculation. A rat IgG isotype antibody was applied as a control. Two-way ANOVA with Tukey’s multiple comparison test. Data are presented as mean values±SEM. The statistical comparisons among these groups were listed as follows: WT+IgG vs KD+IgG, p=0.0258; WT+CTLA4 vs KD+CTLA4, p=0.0994; WT+PD-L1 vs KD+PD-L1, p=0.0016; WT+CTLA4 + PD-L1 vs KD+CTLA4 + PD-L1, p=0.0412; KD+IgG vs KD+CTLA4, p=0.0016; KD+IgG vs KD+PD-L1, p=0.0177; KD+CTLA4 vs KD+CTLA4 + PD-L1, p=0.0066; KD+PD-L1 vs KD+CTLA4 + PD-L1, p<0.0001. (c) Individual tumor growth curves and percentages in different groups of mice. (d) Tumor growth curve in mice with cured tumors after subcutaneous reimplantation of HNSCC. Two-way ANOVA with Tukey’s multiple comparison test was performed. Data are presented as mean values±SD. (e) Expression levels of immune markers and cytokines in tumors of mice in different groups. Kruskal-Wallis with Dunn’s multiple comparison test are reported. ANOVA, analysis of variance; ICIs, immune checkpoint inhibitors.