Figure 6

Inhibition of Dicer prevents HaCaT cell migration. (A) At ∼40% confluence, HaCaT cells were transfected with the scramble (SCR) or Dicer siRNA (10–100 nmol/L) and, after 24 h, total RNA was isolated and the levels of Dicer mRNA was quantified by qRT-PCR, as described in Materials and Methods. 18S rRNA was used as the endogenous control. (B) Cells transfected as in (A) were lysed, and lysates (90 µg) were resolved on SDS-PAGE and subjected to Western blot analyses using anti-Dicer antibody. Lamin was taken as the loading control and densitometric analyses of the same are given in the panel below the Western blots. (C) HaCaT cells were transfected with either the scramble or Dicer siRNA and, after 24 h, a wound was made on the cell layer with a pipette tip. The movement of cells across the wound area at 6, 12 and 24 h was observed and photographed (10×). (D) The wound area at each time point in (C) was quantified. All experiments were in triplicate, and values are means ± SEM of three sets of independent observations. **p < 0.01 and ***p < 0.001 compared with the values in the presence of scramble at the same time points.