Fig. 4.

GRP78-induced M2 macrophage polarization is dependent on JAK/STAT activation. A, B PBMC-derived macrophages were transfected with the indicated shRNAs and then cultured for the indicated time in the presence of 20 ng/mL IL-4 and 20 ng/mL IL-13. The phosphorylation and total content of STAT6 and STAT3 (A) and JAK1 and JAK2 (B) were measured by western blotting. β-actin was used as the loading control. N = 6. C, D Macrophages were transfected with the indicated plasmid and then cultured for 24 h or 48 h without or with the JAK1/2 inhibitor ruxolitinib at 100 nM or the JAK1 inhibitor AG490 at 10 μM. C The polarization of M2 macrophages was analysed by FACS. N = 6. D Statistical analysis of the proportions of M2 macrophages in C. E PBMC-derived macrophages were transfected with the empty vector or GRP78-overexpressing plasmid and then cultured in the presence of 20 ng/mL IL-4 and 20 ng/mL IL-13 for 48 h. The expression of NOTCH1 and HES1 and the phosphorylation and total content of AKT and ERK were measured by western blotting, and GAPDH was used as the loading control. N = 6 A–E The results are representative of three independent experiments. D, E Error bars represent the mean ± SD. p values were determined by one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. ***p < 0.001, **p < 0.01, *p < 0.05