Fig. 3

Autophagy promotes human hair follicle cycle and HFSC activation. A Representative immunofluorescence images of human hair follicles treated with control,3-MA(8 h) or Rapa(8 h) stained with anti-LC3B antibody(red) and anti-K15 antibody(green). B Representative immunofluorescence images of hair follicles treated with control,3-MA(8 h) or Rapa(8 h) stained with anti-P62 antibody(green) and anti-K15 antibody(red). C Quantification of LC3B-fluorescent dots/cell in K15 + HFSCs region of control, 3-MA-treated or Rapa-treated hair follicles(left); Quantification of P62-fluorescent signal in K15 + HFSCs region of control,3-MA-treated or Rapa-treated hair follicles(right). D Human hair follicles were treated with control,3-MA or Rapa. Photographs were taken for 7 consecutive days. E Increased hair shaft elongation after Rapa treatment. Independent experiments were repeated at least three times and data were based on 30 hair follicles from three donors. F Recording of hair follicle cycle stages treated with control,3-MA or Rapa on day 7. G, H Rapa promotes HFSC activation. G The proliferative HFSCs were assessed by the proliferative marker, Ki67 (green) on post-treatment day 3. H Quantification of Ki67 + cell numbers in control,3-MA-treated or Rapa-treated hair follicles. I Representative immunofluorescence images of human hair follicles treated with control,3-MA or Rapa stained with anti-K15 antibody(green) on 12 h and 72 h post-treatment. The data represent the means ± S.E.M. from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, determined by Student’s t-test, 3-MA or Rapa versus Control