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. 2014 Mar 13;289(18):12680–12692. doi: 10.1074/jbc.M114.558981

FIGURE 7.

FIGURE 7.

Enhancement of TGF-β-mediated biological responses. A, decrease in C18ORF1 expression. A549 cells stably expressing C18ORF1 shRNA#1 or shRNA#2 were established. The expression of C18ORF1 and β-actin was detected by RT-PCR. B, increased motility in C18ORF1 knockdown cells. A549 cells carrying C18ORF1 shRNA#1-expressing, shRNA#2-expressing, or control vector were confluently seeded in 24-well plates. After cell scratching, TGF-β was added to the media. Then cell movement was measured by microscopy. The velocity of cell movement was calculated. The experiments were carried out twice. All values represent mean ± S.D. Significantly different from mock in the presence of TGF-β: **, p < 0.01; ***, p < 0.001. C, enhancement of TGF-β-mediated EMT in C18ORF1 shRNA-expressing A549 cells. The cells carrying C18ORF1 shRNA#1, C18ORF1 shRNA#2, or control shRNA were stimulated with or without TGF-β for 36 h. The actin stress fibers were visualized using phalloidin. D, expression of E-cadherin in A549 cells carrying C18ORF1 shRNAs. The cells stably transfected with C18ORF1 shRNA#1, C18ORF1 shRNA#2, or control shRNA were stimulated with TGF-β for indicated times. Then the expression of E-cadherin (upper panel) or β-actin (lower panel) was observed. E, immunofluorescence staining of E-cadherin in cells. The cells carrying C18ORF1 shRNA#1, C18ORF1 shRNA#2, or control shRNA were fixed and then stained with anti-E-cadherin antibody (red). Nuclei were stained blue. Cells were stimulated with TGF-β for 36 h. WB, Western blot.