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. 2022 Jul 2;70(11):2079–2092. doi: 10.1002/glia.24239

FIGURE 2.

FIGURE 2

TRAP1/Smad signaling is activated in primary mouse astrocytes by IL‐17. Primary mouse astrocytes were cultured in a serum‐free medium overnight, prior to stimulating with IL‐17 (50 ng/ml) at indicated time point. (a) The mRNA levels of TRAP1 and TGFβRII were determined by real‐time PCR assay. (b) The protein levels of TRAP1, TGFβRII, Smad4, p‐Smad2/3, Smad2/3, and GFAP in astrocytes stimulated with IL‐17 were measured by Western blot assay. (c) Co‐immunoprecipitation (co‐IP) assay was employed to measure the interaction of TRAP1 with Smad4 in astrocytes treated with IL‐17. (d) The mRNA levels of TGF‐β, TNF‐α, CXCL10, and MCP‐1 in astrocytes with IL‐17 were measured by real‐time PCR assay. (e) The secretion levels of TGF‐β, TNF‐α, CXCL10, and MCP‐1 were measured by ELISA in the supernatant from astrocytes treated with IL‐17. The data are from three independent experiments and represented as the mean ± SEM. *p < .05, **p < .01, and ***p < .001 versus 0 h group treated with IL‐17 (n = 3)