FIGURE 2.

Establishment of DUSP7 knockdown and stable overexpression in SIHA cells. Stable DUSP7‐overexpressing cell lines were established by transducing SIHA cells with Lv‐DUSP7 and were named DUSP7‐SIHA cells (Figure 2A,B; optical magnification*20). The qRT‐PCR results showed that the DUSP7 mRNA level in DUSP7‐SIHA cells was significantly higher than that in NC‐SIHA cells (42.52 vs. 1, p < 0.0001; Figure 2C). Three specific shRNAs targeting DUSP7 were designed and synthesized. The qRT‐PCR results showed that the silencing efficiencies of these shRNAs were 55%, 54% and 47% (all p < 0.0001) when shNC was used as a reference. Cells infected with the most effective (55%) shRNA sequence (GCAUCAAGUAUAUCCUCAATT) were named shDUSP7‐SIHA and used for subsequent experiments. The WB results indicated that the DUSP7 expression level in Dusp7‐SIHA cells was significantly higher than that in NC‐SIHA and wild‐type SIHA cells. In contrast, DUSP7 expression was significantly downregulated in shDUSP7‐SIHA cells compared with Dusp7‐SIHA and wild‐type SIHA cells (Figure 2D,E). *** p < 0.001, **** p < 0.0001