Granzyme B/H Antibody - #AF0175

Fig. 1: PHF8 loss induces a growth-inhibiting immune response. a–d Tumor growth curves a, c and Kaplan-Meier survival curves b, d of Phf8 KO CT26 and MC38 cells and their corresponding vector control cells in immunodeficient mice. In each cases, about 500,000 (CT26) or 1,000,000 (MC38) tumor cells were inoculated subcutaneously into nude mice, and tumor formation was monitored. n = 7 tumors per group. e–h Tumor growth curves e, g and Kaplan-Meier survival curves f, h of Phf8 KO CT26 and MC38 cells and their corresponding vector control cells in syngeneic immunocompetent mice. n = 7 tumors per group. i, j Tumor growth curves i and Kaplan-Meier survival curves j of Balb/c mice subcutaneously injected with about 500,000 the vector control or Phf8 (exogenously transduced pLVX-Phf8 vector) CT26 cells. n = 6 tumors per group. k, l tumor growth curves k and Kaplan-Meier survival curves l of host mice after rechallenge with 1,000,000 Phf8 wild-type CT26 cells in Balb/c mice that remained tumor-free for 38 days after initial challenge with 500,000 Phf8 KO CT26 cells. Control referred to tumor-naive Balb/c mice challenged with Phf8 wild-type CT26 cells. n = 7 tumors per group. m Immunofluorescence staining (left) and quantitative data (right) of CD8+ (CD8+ T cells), granzyme B+ (GZMB+), interferon-γ+ (IFN-γ+) in the vector control and shPhf8 CT26 tumors. n = 5 biologically independent samples. Fluorescent density for each sample was analyzed. Immunohistochemical images shows Phf8 knockdown efficiency. Scale bar, 50 µm. n, o Tumor growth curves n and Kaplan-Meier survival curves o of Balb/c mice injected with 500,000 Phf8 wild-type CT26 cells that were treated with IgG control or anti-PD-1 antibodies, or the same number of shPhf8 CT26 cells treated with IgG control or anti-PD-1 antibodies. IgG or anti-PD-1 treatment began at day 8. n = 7 tumors per group. Data are presented as the mean ± sem. TF, tumor free. Two-way ANOVA in e, g, i, k, and n, log-rank test in f, h, l, and o, and unpaired two-sided Student’s t-test in m. Source data are provided as a Source Data file.

Figure 6. (A) Immunofluorescent CLSM images of B16F10 tumor slices at the end of treatment, NK cells were identified by the antimouse NKp46 antibody (red), and nuclei were visualized by DAPI (blue). Scale bar: 100 μm. (B) Flow cytometric analysis of intratumor NK cell populations identified by PE-conjugated antimouse NKp46. Error bars indicate SD (n = 3). ** and *** represent p < 0.01 and p < 0.001, respectively. (C) Typical immunohistochemical images of the B16F10 tumor sections stained with antibodies against IL-2 and GZMB. Scale bar: 100 μm. (D) Levels of IL-2, GZMB, and IFN-γ in tumor and (E) plasma analyzed by ELISA kits at the end of treatment. Error bars indicate SD (n = 3), *, **, and *** represent p < 0.05, p < 0.01, and p < 0.001, respectively. (F) Images of the expression level of intratumor phosphorylated Smad3 (pSmad3) and Smad3 by western blotting analysis, (a) PBS, (b) M-D@DOX, (c) free DOX/SIS3, (d) M-N@SIS3, (e) M@DOX/SIS3, and (f) MDN@DOX/SIS3. (G) Semiquantitative results of the expression levels of p-Smad3 and Smad3. Error bars indicate SD (n = 3), ** and *** represent p < 0.01 and p < 0.001, respectively.

Fig. 3. FP elicits activated T-cell infiltration in tumours. MC38 tumour-bearing mice were treated with FP or anti-Ly6G+FP. A. Absolute numbers of cells per 1.0 mg of tumour tissue were calculated in dissociated tumours by flow cytometry analysis. B. The percentages of CD69+CD8+/CD3+ and CD69+CD4+/CD3+ cells. C. Representative images of anti-CD4, anti-CD8, or anti-granzyme B/H IHC of tumours and the calculation score are shown. Scale bars, 100 μm. Unpaired two-tailed Student’s t test was performed between two groups, and one-way ANOVA was performed among different groups. Data are shown as the mean ± SEM from at least three independent experiments: *p＜0.05.

FIGURE 6 Baicalein inhibits tumor growth and PD‐L1 expression in vivo. (A) Different nutrient calorie percentages of HFD and CD. (B) Schematic diagram of obesity modeling and subsequent intervention process. (C) ELISA detects serum leptin in HFD and CD mice. (D, E) Body weight growth curve and body weight difference for mice fed with CD or HFD. (F) Tumor growth curve and tumor size diagram for each subgroup, N = 6/subgroup. (G) Immunohistochemistry shows the expression of CD8, PD‐L1, LEP, GZMB, p‐STAT3, and SREBP1 in each subgroup. Scale bar = 100 μm. (H) H score for CD8, PD‐L1, LEP, GZMB, p‐STAT3, and SREBP1 in each subgroup and the statistical analysis of differences in each subgroup. (I) Schematic diagram of baicalein in inhibiting breast cancer (BC) by regulating the tumor microenvironment (Data are shown as the mean ± SD values. Significance was calculated using Student's t test.