Extended Data Fig. 6 | shRNA-mediated knockdown of MCT1 inhibits
melanoma metastasis in vivo. Related to Fig. 2. a, b, Western blot analysis of
MCT1 (a) or MCT4 (b) in subcutaneous tumours from mice xenografted with
efficiently metastasizing melanomas transfected with scrambled control
shRNA or with two different shRNAs (1 and 2) against MCT1 (a) or M C T4 (b).
Wild-type HCC15 cells were used as a positive control for MCT1 (WT, a) and
MCT4 (WT, b) and MCT1-deficient (KO, a) or M C T4-deficient (KO, b) HCC15 cells
were used as a negative control (representative of two independent
experiments). c–e, Growth of subcutaneous tumours (c) in mice transplanted
with melanomas transfected with scrambled control shRNA or shRNAs (sh1 and
sh2) against MCT1. The number of mice analysed in each treatment is indicated
in (one experiment per melanoma). The frequency of circulating melanoma
cells in the blood (d) and metastatic disease burden based on bioluminescence
imaging (e) in the same mice were determined. f, Western blot analysis of MCT1
in subcutaneous tumours transfected with scrambled control shRNA or
shRNAs against MCT1, with (MCT1-OE) or without an shRNA-insensitive MCT1
cDNA. g, h, Growth of subcutaneous tumours (g) and metastatic disease
burden at end point (h) in mice transplanted with melanomas transfected with
scrambled control shRNA or shRNAs against MCT1 and an shRNA-insensitive
MCT1 cDNA. i, Fold change in mean f luorescence intensity for CellRox DeepRed
staining (ROS) in xenografted melanoma cells with scrambled control shRNA
or shRNAs against MCT1 treated with AZD3965 or DMSO. Data are mean ± s.d..
Statistical significance was assessed using nparLD followed by Benjamini–
Hochberg’s multiple comparisons adjustment (c), log 2 -transformed one-way
ANOVAs with Holm–Sidak’s multiple comparisons adjustment (d, e, h), mixed-
effects analysis followed by Dunnett’s multiple comparisons adjustment (g) or
log 2 -transformed two-way ANOVA with Sidak’s multiple comparisons
adjustment (i).