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. 2014 Jan 21;9(1):e85906. doi: 10.1371/journal.pone.0085906

Figure 3. Effect of Sfrp5 on insulin signaling in primary human adipocytes.

Figure 3

Primary human adipocytes were kept untreated, exposed to Sfrp5 for 24/ml TNFα for 24 h with or without a 4 h preincubation with Sfrp5. Then, when indicated (+) cells were stimulated with insulin (10 min; 100 nM). Cell lysates were analyzed for phosphorylation of Akt-Thr308 (A), Akt-Ser473 (B), GSK3α-Ser21 (C) and PRAS40-Thr246 (D) by Western blotting. Phosphorylation signals were normalized for GAPDH protein abundance and expressed as mean ± standard error of the mean of five independent experiments using cells from different donors. The values obtained for Sfrp5-untreated, insulin-treated cells were considered as control and set at 100%. Differences among groups were calculated by two-way ANOVA followed by Bonferroni multiple comparison analysis. #, indicates p<0.05 for the effect of insulin stimulation; ***, p<0.001; **, p<0.01; *, p<0.05 versus untreated cells.