Fig. 8 | Nature Communications

Fig. 8

From: Transcriptional regulation of autophagy-lysosomal function in BRAF-driven melanoma progression and chemoresistance

Fig. 8

TFEB S142 phosphorylation confers BRAFi resistance in melanoma cells. a Representative images (left) and quantification (right) of the colonogenic survival of A375 cells stably expressing control shRNA (pGIPZ) or TFEB shRNA complemented with empty vector, WT TFEB, and S142A or S142E TFEB mutants that are shRNA-resistant, after treatment with DMSO or PLX4720 at the indicated concentrations. Endogenous and reconstituted TFEB expression was confirmed by immunoblotting (bottom right). n= 3 independent experiments. b Bioluminescence images (top) of tumor regression of the indicated A375R xenograft tumor genotype in live NOD/SCID mice at the indicated time after inoculation. Radiant efficiency expressed as p/s/cm2/sr/(μW/cm2) was quantified (bottom). c Effect of PLX4720 on tumor response of xenografts formed by the indicated A375 cell lines. After tumor establishment (~ 500 mm3), mice bearing A375 xenografts were treated with PLX4720 (20 mg/kg, i.p.) daily for 21 days. Values are the mean tumor volume ± SD per time point for 6–7 mice per group. d, e Representative gross images (d) of livers with metastatic nodules (top), H&E-stained sections (second row) and IHC analysis of TGF-β and p-Smad3 of the indicated A375 xenograft genotypes 21 days after PLX4720 treatment (20 mg/kg, i.p., daily). Scale bars, 100 μm. The numbers of metastatic nodules in the liver in (d) was quantified in (e) (n= 5–6 mice per group). For all quantification, data represent the mean ± SD derived from indicated number of independent experiments. Comparisons were made using Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.001; n.s. not significant

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