Fig. 2: Caveolin-1 depletion in stimulated melanocytes increases cAMP production and pigmentation.
From: Coupling of melanocyte signaling and mechanics by caveolae is required for human skin pigmentation
a, b Quantification of intracellular cAMP fold-change in melanocytes. a Melanocytes were transfected with Ctrl (control) or Cav1 (caveolin-1) siRNA for 24 h and incubated with DMSO (dimethylsulfoxide) or 30 µM of FSK (forskolin) for 3 h. b Melanocytes were treated with Ctrl (scrambled) or CavTratin (Cav1- scaffolding domain) peptides for 7 h and incubated with DMSO or 30 µM of FSK for 1 h. c Quantification of TYR (Tyrosinase) mRNA expression in siCtrl- or siCav1-treated cells maintained in supplemented medium or in poor medium + FSK (3 h) and normalized to GAPDH (glyceraldehyde 3-phosphate dehydrogenase). d–g Melanocytes treated 5 days with siCtrl or siCav1. d Immunoblot analysis of melanocytes treated for 5 days with siCtrl or siCav1 using the indicated antibodies (left) and associated quantifications (right). e Estimation of intracellular melanin content normalized to control. f Conventional EM images representative of each condition with the respective zooms of the boxed regions; Bar: original 1 µm, zoomed 0.5 µm; II to IV represent different stages of maturation of melanosomes. g Quantification of the number of non-pigmented and pigmented melanosomes from EM images as in f (n = 14 independent cells). Values are mean ± s.e.m. a Five independent experiments; b, c three independent experiments; d, e four independent experiments. g 19 cells, 4 independent experiments. a, g One-way ANOVA with Holm–Sidak’s multiple-comparison test; b, c, e two-tailed unpaired t-test with Welch’s correction; d two-tailed paired t-test. See also Supplementary Table 2.
