Fig. 6: DP cell condensation activity in keratin-eliminated conditioned medium of TGFβ2-treated ORS cell culture using a column containing anti-human type I + II hair keratin antibody-conjugated beads, and germ formation in ORS cell culture and KRT31/KRT34 knockdown ORS cell culture in the presence of TGF-β2.
From: Keratin-mediated hair growth and its underlying biological mechanism
a Western blot image of the keratin-eliminated conditioned medium and the keratin-bound beads. b Graphical quantification of growth factor content of the conditioned medium and the keratin-eliminated conditioned medium collected from TGFβ2-treated ORS cell culture using antibody array for growth factors. c DP cell condensation activity in keratin-eliminated conditioned medium of TGFβ2-treated ORS cell culture using a column containing anti-human type I + II hair keratin antibody-conjugated beads. Immunofluorescent image; phalloidin, red; β-catenin, green; DAPI, blue; DP media, DP culture medium; TGFβ2-ORS CM, conditioned medium collected from TGFβ2-treated ORS cell culture; TGFβ2-ORS CM (IgG-column), conditioned medium collected from TGFβ2-treated ORS cell culture and then treated with normal IgG-conjugated beads; 1% Keratin, DP medium containing 1(w/v)% keratin; TGFβ2-ORS CM (keratin Ab-column), keratin-eliminated conditioned medium collected from TGFβ2-treated ORS cell culture. Graphical representation of DP cell condensation activity. *P,0.05 and **P,0.01, indicate a difference between DP media and experimental groups. #P,0.01, indicates a difference between TGFβ2-ORS CM (IgG-column) and TGFβ2-ORS CM (keratin Ab-column). Scale bars, 100 μm. (n = 4; mean ± standard deviation (s.d.)). d Germ formation of ORS cells and KRT31/KRT34 knockdown ORS cells. Western blot image of keratin content of conditioned media and cell lysates from negative control siRNA-transfected ORS cell culture in the presence of TGFβ2 and KRT31/KRT34 knockdown ORS cells culture in the presence of TGFβ2. Image of ORS cells and KRT31/KRT34 knockdown ORS cells in the presence of TGFβ2. Germ formation of ORS cells and KRT31/KRT34 knockdown ORS cells in the presence of TGFβ2 by immunofluorescent staining; phalloidin, red; P-cadherin, RUNX1, KRT34, green; DAPI, blue. Scale bars, 200 μm.
