Fig. 2

PKMYT1AR knockdown inhibits tumor cell growth and migration. a Establishment of PKMYT1AR overexpression and knockdown cell lines in A549 verified by Real-time RT-PCR. b-d PKMYT1AR knockdown dramatically inhibited A549 cell proliferation (b) and colony formation ability (c), (d) is the quantification data for (c). e-f. Effect of PKMYT1AR knockdown on the G0/G1 cell cycle transition was tested in A549 cells by PI staining and flow cytometry. f is the quantification data for (e). g PKMYT1AR knockdown regulated the expressions of cell cycle transition mediators, including CDK2, CDK6, cyclin D1, p21 and p27. Indicated cell extracts were probed with indicated antibodies. h-i Knockdown of PKMYT1AR inhibited A549 cell migration using wound healing (h) and transwell (i) assays. Quantification data were also indicated, and the OD₅₇₀ values for trans-well assay were indicated below. Scale bar= 50 μm. j Indicated cell extracts were probed with indicated antibodies to examine the expression patterns of cell migration regulators, including E-cadherin, N-cadherin, Vimentin and Slug. k-m PKMYT1AR knockdown inhibited xenograft tumor formation in vivo. Representative xenograft tumor images (k), tumor masses (l) and tumor volumes (m) were shown. n-o Representative IHC staining of Ki67 (n) and Cleaved Caspase 3 (CC3, o) for indicated xenograft tumors. Quantification data were also indicated. Scale bar=50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001. HPF=high power field, pCDH-Vec=pCDH lenti-viral plasmid vector control. ove=over-expression, sh#1=shRNA#1, sh#2=shRNA#2