Fig. 4

DDA induces autophagy in melanoma in vivo and in vitro in an LXRβ-dependent manner. a, b Mice engrafted with human SKMEL-28 or mouse B16F10 cells (10 per group) were treated with DDA (i.p. 20 mg/kg/day) or vehicle. Mean tumor volumes ± S.E.M. are shown, **P < 0.01, analysis of variance (ANOVA). Data are representative of three independent experiments. At the end of treatments, tumors were analyzed for Nur77, NOR1 and LC3 protein expression by a, b immunoblotting or c immunohistochemistry (brown staining). c Representative immunohistochemical analysis. Magnification ×40. Insert is 4× digital amplification. d, e TEM images of cells transfected with shCTRL, sh3LXRβ, or sh4LXRβ and treated with control vehicle or 2.5 µM DDA for 24 h. e TEM images of cells transfected with shCTRL, sh3LXRβ, or sh4LXRβ and treated with 2.5 µM DDA for 24 h. d, e N, nucleus; C, cytoplasm; AM, amphisome