Fig. 2.

Activation of α7 nAChR enhances phosphorylation of p38 MAPK by downregulatingDUSP1andDUSP6. (A) MSigDB canonical pathways enrichment analysis of differentially expressed genes (DEGs) (P < 0.05) after GTS-21 treatment, bubble size indicates the absolute gene counts enriched in a term. (B) Volcano plot of DEGs comparing GTS-21-treated versus untreated (medium) cells. Three samples in either medium or GTS-21 treated ACH2 groups were used for RNAseq analysis. (C) ACH2 were treated with GTS-21 (40 μM) and detect the phosphorylation of MAPKs family members p38 MAPK, JNK and ERK, and gag protein level by western blotting. (D) The endogenous DUSP1/6 was knocked-down with lentiviruses containing specific shRNAs. (E) The gag mRNA level of negative control and DUSP1+6 knockdown groups were identified by RT-qPCR. (F) The levels of gag and p-p38 MAPK in two groups were detected by western blotting. (G) The inhibitor of DUSP1/6 were used to inhibit DUSP1 and DUSP6 for 8 ​h, then the gag and p-p38 MAPK levels were detected by western blotting. Result is one representative from three independent repeats. Data are presented as mean ​± ​SD. ∗∗P ​< ​0.01, ∗∗∗P ​< ​0.001 and ∗∗∗∗P ​< ​0.0001 denote significant difference.