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. 2015 Sep 4;4(10):1213–1221. doi: 10.1242/bio.011973

Fig. 4.

Fig. 4.

Acceleration of melanosome generation by ET-1 counteracting autophagy-driven degradation in melanocytes. (A) NHEMs were treated with 10 nM ET-1 for the indicated times. Cells were harvested for western blotting analysis using LC3-, p62- or Pmel17-specific antibodies. The blots were then re-probed with a β-actin-specific antibody for a loading control. (B) Relative intensity of each band normalized to β-actin was expressed as a ratio against the control. Values represent means±s.d. from three different experiments. **P<0.01; *P<0.05 (ANOVA, Holm test). (C) NHEMs were cultured with or without 10 nM ET-1 for 24 h and were then subjected to immunofluorescence staining with an anti-p62 antibody (green) and an anti-Pmel17 antibody (red). Merged images with nuclear staining (DAPI) are shown. The colocalization of p62 and Pmel17 is observed as yellow dots. Scale bar=50 µm. (D) The values of colocalization of p62 and Pmel17 were quantified and expressed as Pearson's correlation coefficient by ImageJ analysis software. Values represent means±s.d. from four samples. *P<0.05 (Student's t-test).

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