Figure 4.

miR-184 Repressed the Clonogenic Potential and Accelerated Epidermal Differentiation

(A and B) Primary human foreskin KCs were transfected with pre-miR-184-mimic (PM184) or anti-miR antagonist (AM184) or appropriate controls (Ctl-PM and Ctl-AM, respectively) and then subjected to the clonogenicity test. Colonies were visualized by Rhodamine staining (A, examples of two experiments are shown) and quantification by computerized analysis is shown in (B).

(C and D) Primary human KCs were induced to differentiate/stratify by high calcium, and quantitative real-time PCR analysis of the indicated genes was performed to validate differentiation efficiency (C) and TaqMan assay to test the expression of miR-184 (D).

(E) Primary KCs were transfected with PM184 mimic, or with disease-causing pre-miR-184-mutant (C57U) mimic or antagonist (AM184), or the appropriate control oligonucleotides, and then subjected to calcium-induced stratification/differentiation. Real-time PCR analysis was performed to evaluate the effect of transfection on differentiation.

Data shown are means ± SD from three independent experiments. ^∗p < 0.05 statistically significant by Student's t test.