FIGURE 8.

PMEL cysteine mutants traffic appropriately to late endosomal compartments when expressed in HeLa cells. HeLa cells transiently transfected with wild-type PMEL (a–c), PMEL C60S (d–f), PMEL C130S (g–i), C138S (j–l), C301S (m–o), C475S (p–r), C516S (s–u), C525S (v–x), C533S (y, z, and aa), C541S (ab–ad), C550S (ae–ag), or C566S (ah–aj) were fixed, labeled with antibodies to PMEL (NKI-beteb, red: a, d, g, j, m, p, s, v, y, ab, ae, and ah) and LAMP1 (H4A3, green: b, e, h, k, n, q, t, w, z, ac, af, and ai), and analyzed by deconvolution immunofluorescence microscopy. Shown are representative images of each label separately and merged together (c, f, i, l, o, r, u, x, aa, ad, ag, and aj). Arrows indicate examples of overlap between PMEL and LAMP1. Insets show ×4 magnification of the boxed regions. Scale bar represents 10 μm.