Fig 1: Effects of EGLN1 and EGLN3 on dermal papilla cells (DPCs) and hair follicles (HFs). (a) mRNA expression levels of EGLN1 and EGLN3 were significantly downregulated after corresponding small interfering (si)RNA transfection in cultured DPCs. (b) xCELLigence system detection showed that siEGLN1 or siEGLN3 treatment significantly increased the proliferation of DPCs compared with the negative control (NC) group DPCs. (c) Immunofluorescence staining assay identified decreased protein levels of EGLN1 and EGLN3 after siEGLN1 or siEGLN3 treatment of HFs. (d) EGLN1 or EGLN3 knockdown increased the length of hair shafts compared with the NC after 2 days of culture. (e) Macroscopic images of cultured hair follicles were obtained every other day. The hair cycle stage was macroscopically determined on day 6, and a greater percentage of anagen HFs and a lower percentage of catagen HFs remained in the EGLN1 or EGLN3 siRNA‐treated group compared with the NC group. Data are expressed as the mean (SD) of each group. P‐values were calculated using unpaired‐sample t‐tests. ***P < 0·001, **P < 0·01, *P < 0·05. [Colour figure can be viewed at wileyonlinelibrary.com]
Fig 2: Validation of the expression of differentially expressed genes involved in the hypoxia‐inducible factor (HIF)‐1 and Wnt signalling pathways. (a) Haematoxylin and eosin staining showed that the hair follicles (HFs) were anagen HFs and the bulb diameter was decreased in vertex HFs compared with occipital HFs. (b) The expression of HIF‐1 pathway‐related genes (EGLN1, EGLN3 and HMOX1) and Wnt signalling pathway‐related genes (PEDF/SERPINF1, SFRP2 and LGR5) was examined in vertex HFs compared with occipital HFs. The mRNA expression of EGLN1, EGLN3, PEDF/SERPINF1 and SFRP2 was significantly higher in the vertex HFs compared by paired‐sample t‐tests. (c) Immunofluorescence staining showed increased protein expression of EGLN1, EGLN3, PEDF or SFRP2 in vertex HFs compared by paired‐sample t‐tests. **P < 0·01, *P < 0·05. [Colour figure can be viewed at wileyonlinelibrary.com]
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