Fig. 4

JMJD6 Promotes the Proliferation and Invasion of Melanoma Cells in Vitro and in Vivo. a A375 cells were infected with lentiviruses carrying control shRNA or JMJD6 shRNA, and/or infected with retroviruses carrying vector or expression plasmids for PAK1, PAK1Δ15, JMJD6, or JMJD6m. Cells were maintained in DMEM medium for 14 days before staining with 0.5% crystal violet and counting for colony numbers. b A375 cells were infected with viruses carrying indicated constructs, and the cell viability rate was evaluated using Cell Counting Kit-8. c A375 cells were infected with control shRNA/vector, JMJD6 shRNAs, JMJD6, or JMJD6m, and the expressions of epithelial and mesenchymal markers were tested by Western blotting. d The SK-MEL-1 cells infected with viruses carrying indicated constructs were starved for 18 h before cell invasion assays were performed using Matrigel transwell filters. The invaded cells were stained by 0.5% crystal violet and counted under a light microscope. e B16F10 cells were infected with viruses carrying control shRNA or Jmjd6 shRNAs. These cells were subcutaneously injected into the right flank of 6-week-old female C57BL/6 mice (n = 6). The bioluminescence imaging was used to quantify tumor size, and representative in vivo bioluminescent images are shown. f B16F10 cells were injected intravenously through the tail vein of 6-week-old female C57BL/6 mice (n = 6). Lung metastasis was quantified by bioluminescence imaging. Scale bar = 250 μm. Representative in vivo bioluminescent images are shown. Lung cancer specimens were examined by in vitro bioluminescent measurement and stained with hematoxylin and eosin (H&E)