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. 2014 Aug 8;124(9):3863–3878. doi: 10.1172/JCI72189

Figure 9. Resolving MΦs mediate the protective role of the VEGF-C/VEGFR3 pathway during chronic experimental colitis in a STAT6-dependent manner.

Figure 9

(A) Mice undergoing 2 cycles of DSS treatment were injected with Ad-hVEGF-C or anti-VEGFR3 Ab. Colonic expression of p-STAT6 and total STAT6 was evaluated by Western blotting in MΦs after 4 hours in culture (in vivo) and in BM-derived differentiated MΦs (in vitro). Actin was used as a loading control. (B and C) BM-derived MΦs were cultured, transfected with a STAT6-specific siRNA or a STAT6 scramble, and stimulated in vitro with VEGF-C for 24 hours. Expression of the indicated genes was then examined by ELISA (B) or real-time PCR (C). Data represent the means ± SEM of 3 independent experiments. **P < 0.01. (D–F) GFP plus BM-derived MΦs (BMDMs) were cultured, transfected with a STAT6-specific siRNA or a STAT6 scramble, and stimulated in vitro with VEGF-C or mF431C1 (D). Cells were then injected into the rectal mucosa of DSS-treated mice (n = 4/group), and clinical parameters such as body weight (E) and DAI (F) were evaluated. Black dashed lines represent the 2 DSS cycles. Results are presented as the mean value per group at the indicated time points ± SEM. *P < 0.05 for DSS plus VEGF-C MΦs versus DSS alone; §P < 0.01 for DSS plus VEGF-C plus STAT6 siRNA MΦs versus DSS plus VEGF-C MΦs.