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. 2023 Oct 16;133(20):e161366. doi: 10.1172/JCI161366

Figure 2. VEGFR2 pY1173/PLCγ signaling in experimental tumors affects vascular leakage, response to therapy, and antitumor immunity.

Figure 2

(A and B) Immunostaining (A) and mean fluorescent intensity (MFI) quantification (B) of pPLCγ Y783 in Vegfr2+/+ (WT) and Vegfr2Y1173F/+ (Y1173F/+) B16F10 melanoma tumors; n = 5 (WT) and 3 (Y1173F/+) mice, ≥ 3 fields of view/experiment. Scale bar: 100 μm. Unpaired 2-tailed Students’ t test. (C) Tumor growth of WT and Y1173F/+ B16F10 tumors; n = 5 (WT) and 3 (Y1173F/+) mice. 1-way ANOVA. (D) Quantification of vessel density (MFI) from A; n = 5 (WT) and 3 (Y1173F/+) mice, ≥ 3 fields of view/tumor. Unpaired 2-tailed Students’ t test. (E and F) Representative image (E) and quantification (F) of extravasated fixable 2,000 kDa FITC-dextran in B16F10 tumors; n = 5 (WT) and 3 (Y1173F/+) mice, ≥ 3 fields of view/experiment. Scale bar:100 μm. Unpaired 2-tailed Students’ t test. (G) Fluorescent intensity of 70 kDa TRITC-dextran extracted from B16F10 tumors; n = 5 (WT) and 3 (Y1173F/+) mice. Unpaired 2-tailed Students’ t test. (H and I) Tumor growth (H) and tumor weights (I) of WT and Y1173F/+ B16F10 tumors treated with 5 mg/kg Temozolomide (TMZ) or DMSO control; n = 9–10 mice/group. 2-way ANOVA (H), 1-way ANOVA (I). (J–M) Percent B cells (CD19) (J), helper T cells (CD3 and CD4 costaining) (K), regulatory T cells (CD3, CD4, CD25, and Foxp3 costaining) (L), cytotoxic T cells (CD3 and CD8 costaining) (M) in WT and Y1173F/+ B16F10 tumors; n = 16/genotype. Unpaired 2-tailed Students’ t test. (N) mRNA expression of TGFβ (Tgfb1-3), IL10 (Il10), IL35 (Il12a, Ebi3), IFNγ (Ifng), and TNFα (Tnf) in WT and Y1173F/+ B16F10 tumors; n = 5 mice/genotype. Unpaired 2-tailed Students’ t test. (O) Tumor weight of CD40- or IgG-treated WT and Y1173F/+ B16F10 tumors harvested at day 12; n = 9–10 mice/group. 1-way ANOVA. Data represent mean ± SD or SEM (H). *P < 0.05, **P < 0.01, ***P < 0.001.