MiR-3661 specifically targets PPP2CA expression in human cells. (A) RNA levels of PPP2CA and PPP2CA-iso were measured from different human cell lines, MRC5SV, Beas2B, A549, and H1299, that were transfected with the control RNA (mock) or miR-3661 mimic at 30 nM final concentration. (B) The protein levels of PP2Acα were measured from MRC5SV, Beas2B, A549, and H1299 cells transfected with the control RNA (mock) or miR-3661 mimic at 30 nM final concentrations. β-actin was used as the internal loading control. (C) The protein levels of PP2Acα were measured from 293FT cells transfected with 30 nM of a control RNA or the predicted miRNAs as indicated. (D) The protein levels of PP2Acα were measured from 293FT cells transfected with the control RNA (mock), miR-3661 mimic alone or together with a miR-3661 inhibitor at 30 nM final concentration. (E) (Left panel) the top sequence is the mutant plasmid construction of PPP2CA, light gray: the mutated sites of the PPP2CA sequence targeted by Hsa-miR-3661 (bottom sequences). (Right panel) The protein levels of Flag-PP2Acα were measured from 293FT cells transfected with 30 nM of wild-type (WT) or mutant (MT) Flag-PP2Acα together with the mock or miR-3661.
