Figure 4

Downregulation of Tyrosinase related protein 1 (TRP1) by selected miRNA at mRNA and protein levels. (a) TRP1 mRNA profiling was done by qPCR and normalized with NC (negative control i.e., scrambled control). Expression of TRP1 in keratinocytes at mRNA level after transfection with pre-miR-326, pre-miR-518a-5p and pre-miR-518c, (b) Transcriptional expression of TRP1 in keratinocytes after transfection with pre-miR-185, pre-miR-202-3p, pre-miR-423-3p and pre-miR-525-5p (c) Western blot of TRP1 expression after transfection with pre-miR-326, pre-miR-518a-5p, pre-miR-518c. UT is Untransfected control. Full gel pictures are shown in Supplementary Fig. 12. (d) Western blot of TRP1 expression after transfection with pre-miR-185, pre-miR-202-3p, pre-miR-423-3p and pre-miR-525-5p. Full gel pictures are shown in Supplementary Fig. 13. (e) Densitometric analysis of TRP1 protein expression in pre-miR-326, pre-miR-518a-5p, pre-miR-518c transfected keratinocytes after normalization with β- Actin, (f) Densitometric analysis of TRP1 protein expression in pre-miR-185, pre-miR-202-3p, pre-miR-423-3p and pre-miR-525-5p transfected keratinocytes after normalization with β-Actin (N = 3), (g) Target validation assay results showing significantly reduced relative luciferase activity of 3'UTR clones transfected with pre-miR-202-3p, pre-miR-518a-5p and pre-miR-525-5p. All the bars showing reduced expression are depicted as mean ± standard error of mean (S.E.M).