Figure 2.

Ψ/m5C Substitution Prevents Increased Silencing by Additional miR-126-3p Target Sites

(A–D) Representative GFP and GAPDH immunoblots (A and C) and densitometric quantification (B and D) of HEK293 cells transfected with miR-126-3p or miR-143-3p mimic or vehicle, 24 h before transfection with the indicated GFP miRNA switch (A and B) or HUVECs transfected with the indicated 100% Ψ/m5C-modified GFP miRNA switches (C and D). GFP and GAPDH expressions were measured after 24 h. Data represent the mean ± SEM, normalized against GAPDH (B and D) and relative to vehicle control (B) or untargeted GFP control (D). *p < 0.05 versus miR-143 control (B); versus GFP-treated cells (D). #p < 0.05 for the indicated comparisons. #TS, the number of miR-126-3p target sites in the 3′ UTR of the miRNA switch; NTC, non-transfected control. Numbers above bars indicate the mean values.