Fig 2: Detection of ZYX in AGA hair follicles and dermal papilla cells. (A) HE staining of hair follicles. Scale bar: 5 μm. (B,C) mRNA levels of ZYX in hair follicles. N = 20. (D) Immunofluorescence examination of ZYX in hair follicle tissues. Scale bar: 5 μm. (E) Cell counting of ZYX-positive dermal papilla cells. N ≥ 3; *P < 0.05, ***P < 0.001 Pregnant group vs. Alendronate group. Control bars represent the mean ± SD.

Fig 3: Proliferation ability and inductivity properties in ZYX-deficient DP cells. (A) ZYX levels in DPCs after NC or si-ZYX treatment. (B,C) RTCA assay of si-ZYX- and NC-treated DP cells. (D,E) Cell cycle distribution in si-ZYX- and NC-treated DP cells. (F) Inductivity marker expression in si-ZYX HFs using qPCR assay. (G) Inductivity ability assay in si-ZYX or control DPCs. *P < 0.05, **P < 0.01, ***P < 0.001. Control bars represent the mean ± SD.

Fig 4: Effect of ZYX on hair growth in mice. (A) Size of hair covered skin area in Zyx-/- and WT mice. Different morphology of hair shaft (B) and hair follicles (C) in Zyx-/- and WT mice. (D) HF percentage and cycling score in Zyx-/- and WT mice. Scale bar: 5 µm, N =6 per group; *P < 0.05, ***P < 0.001. Control bars represent the mean ± SD.

Fig 5: The ratio of hair follicle growth after ZYX knockdown in vitro. (A) Measurement of the length of hair shaft in ZYX knockdown and negative control groups. (B) Distribution of hair follicle cycle in ZYX knockdown and control groups. (C,D) Bulb diameter and cell counting of Ki67-positive cells in si-ZYX HFs and NC HFs. (E) Evaluation of apoptosis using the TUNEL method after ZYX knockdown. (F) Inductivity marker expression in si-ZYX HFs. *P < 0.05, **P < 0.01, ***P < 0.001. Control bars represent the mean ± SD.