Extended Data Fig. 5: Knocking-down CD248 reduced TGF-βRI expression and inhibited the activation of canonical and non-canonical signaling down-streaming of TGF-β receptor.
a, Western-blot analysis of canonical and non-canonical TGF-β signaling pathways (p-Smad2/Smad2, p-Smad3/Smad3, p-ERK/ERK, p-p38/p38) in WT and CD248 KO mouse hearts with or without I/R injury. Statistical analysis was shown in b. n = 6 in each group. c, Western-blot analysis of TGF-βRI and its downstream canonical and non-canonical signaling (p-Smad2/Smad2, p-Smad3/Smad3, p-ERK/ERK, p-p38/p38) in cardiac fibroblasts transfected with scramble or OE-CD248 lentivirus, and statistical analysis was shown in d. n = 3 in each group. e, RT-PCR analysis of TGF-βRI and TGF-βRII mRNA level in CD248-overexpression (OE-CD248) versus scramble-treated cardiac fibroblasts, n = 4 in each group. f, TGF-βRI protein level detected in cardiac fibroblasts treated with Scramble+DMSO, sh-CD248 + DMSO, sh-CD248+baflomycin and sh-CD248 + MG132 separately. Statistical analysis was shown in g. n = 6 in each group. h, Combination analysis of TGF-βRI and CD248 by co-immunoprecipitation in cultured cardiac fibroblasts. CD248 expression analysis in a and f was conducted using the Abcam antibody (no. 67273), while analysis in c and h was performed using the Proteintech antibody (no. 60170). Data are shown as mean ± s.e.m.; unpaired two-tailed Student’s t-test (d and e) and two-way ANOVA (b and g).
