Fig. 4.

RNAi-mediated knockdown of endogenous HsTRPML1 and subsequent rescue of endogenous HsTRPML2 mRNA expression levels. a Real-time QPCR analyses of control (untreated) and RNAi-treated samples. Endogenous HsTRPML1 expression levels were markedly reduced upon shRNA-1208 treatment. A concomitant decrease of HsTRPML2 transcripts was also observed but not with HsTRPML3 transcripts. Data are represented as means ±SEM, N =3 independent trials. *p< 0.05, Student's t test, two-tailed. b Real-time QPCR of endogenous HsTRPML2 transcript levels upon coexpression of pCMV HsTRPML1 with varying concentrations of shRNA-1208 (4μg and 8μg). The decrease of endogenous HsTRPML2 levels upon RNAi treatment was rescued by overexpressing a pCMV-HsTRPML1 construct. Data are represented as means ± SEM, N=3 independent trials. Real-time QPCR reactions were performed as described in the Materials and methods section