Figure 6.

GDF15 regulated DCs by using TGFβ receptors. (A) Expression of GFRAL and TGF-β RI and RII. RNA was extracted from DCs and the expression of GFRAL and TGF-β RI and RII was detected by RT-PCR. PCR products were run on 1.5% agarose gel and visualized with EB. (B) TGF-β receptor I and II reversed the inhibitory function of GDF15 in DC maturation. DCs cultured from WT mice and treated with rhGDF15 on day 4. Ly210976 (5 mg/ml) and Ly 364947 (10 mg/ml) was added to rhgDF15 treated DCs on day 5, respectively. 24 h later, DCs maturation was determined by measuring expression of MHCII, CD40, CD80, CD83, CD86, and CCR7 flow cytometry. Data were representatives of three independent experiments. (C) IP assays. Total protein was extracted from the above DCs treated with GDF15-Ad or control Adenovirus and an IP assay was employed to pull down GDF15 bound protein using GDF15 Abs and control Ig Abs. IP protein was subjected to western blotting with TGFβ RI and RII Abs respectively. Representative of image of n = 4 experiments. PCR products for TGF-β RII from Splenocytes (SP) and two DC samples (DC1 and DC 2) were loaded separately by other genes products on an agarose gel and the image shown was modified by deletion of other genes' bands between TGF-β RII's bands.