PIKfyve regulates melanosome biogenesis
Fig 5
PIKfyve inhibition results in decreased melanin accumulation and decreased accumulation of advanced stage melanosomes.
A) MNT-1 cells were treated with PIKfyve inhibitors YM-201636 (YM), apilimod or vehicle control for five days. A spectrophotometric melanin quantitation assay was used to measure the amount of accumulated melanin. (n = 6 as indicated by error bars). B) Equal numbers of MNT-1 cells treated with the indicated doses of YM or Apilimod were pelleted. Photographs of the cell pellets were obtained to document that YM and Apilimod treatment inhibited the accumulation of melanin. Normal human melanocytes (NHM) were treated with 1000 nM YM-201636 or vehicle without (Control) or with DOPA histochemistry (DOPA Tx) and (C,D) observed by electron microscopy Scale bar, C = 5 μm and D = 2 μm. (E) Pie graphs representing quantification of melanosome stages as percentage in NHM treated with 100, 500, or 1000 nM YM-021636 or vehicle. Each experiment was performed three times in triplicate. For all experiments, data shown are mean ± S.D *, p < 0.05; **, p < 0.01; or ***, p < 0.001 using a Student’s paired T test versus vehicle treated control.