Research Paper Volume 9, Issue 11 pp 2376—2396

An expression screen for aged-dependent microRNAs identifies miR-30a as a key regulator of aging features in human epidermis

Figure 3. Caracterization of reconstructed epidermis overexpressing miR-30a. Reconstructed epidermis (REs) were generated from keratinocytes transduced with the miR-30a lentiviral construction and treated by doxycycline to activate miR-30a expression. (A) Evaluation of the morphology of the REs by HES staining and confirmation of the miR-30a overexpression by in situ hybridization in control and miR-30a RE. (B) Immunofluorescent staining of differentiation markers (LORICRIN, K10, K1, INVOLUCRIN and K14) and immunohistochemical staining of proliferation marker (Ki67) in control and miR-30a overexpressing REs. Counterstaining was performed with DAPI and the polycarbonate membrane is indicated by dotted lines. (C) Quantification of the LORICRIN, K10, K1, INVOLUCRIN and K14 labeled area in the REs and evaluation of the number of KI67-positive cells. Results are mean +/- SD from three independent samples. ****P<0,0001. ns: non-significant. Scale bar = 25 μm. Representative photographs of 3 independent replicates were shown.