Figure 6
Luciferase reporter analysis of miR-141: PTPRG 3′-UTR interaction and siRNA knockdown of PTPRG in PTECs in response to oxidative stress and forced miR-141 expression. (A) RT-qPCR analysis of PTPRG showed significant downregulation in 1 mM H2O2 (50% decrease) and following forced miR-141 expression (30% decrease). (B) PTPRG 3′-UTR reporter assay activity in PTECs showed PTPRG to be a direct miR-141 target. (C) RT-qPCR analysis showed significant PTPRG mRNA knockdown using siRNA, as well as repression of PTPRG in 1 mM H2O2 compared to healthy cells. (D) LDH release increased significantly under oxidative stress and showed a further increase with PTPRG knockdown. (E) Cell viability marker Alamar Blue decreased in 1 mM H2O2, lowering further following PTPRG knockdown. Statistical analysis between groups was carried out using one-way ANOVA with post-hoc analysis of Tukey (n = 3). Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
