Fig. 7

Depletion of PKMYT1AR inhibits cancer stem cell maintenance. a Relative mRNA expressions of PKMYT1AR and cancer stem cell marker genes, including CD44, OCT4, SOX2, Nanog in A549 cells, were examined by Real-time RT-PCR upon PKMYT1AR knockdown. b Tumor sphere formation abilities of indicated cells after PKMYT1AR knockdown were examined. Scale bar=50 μm. c Total extracts of indicated cells were probed with indicated antibodies by immunoblot. d Relative mRNA expressions of PKMYT1 and cancer stem cell marker genes in A549 cells, were examined by Real-time RT-PCR upon PKMYT1 knockdown. e Tumor sphere formation abilities of indicated cells after PKMYT1 knockdown were examined. Scale bar=50 μm. f Total cell extracts of indicated cells were probed with indicated antibodies by immunoblot. g Relative mRNA expressions of PKMYT1 and cancer stem cell marker genes in A549 cells, were examined by Real-time RT-PCR with miR-485-5p mimics or miR-NC co-expression. h-i Tumor sphere formation abilities of indicated cells after miR-485-5p mimics or miRNA controls co-transfection were examined (h) in A549 cells. i is the quantification data for (h). Scale bar=50 μm. j Total extracts of indicated cells were probed with indicated antibodies by immunoblot. k-l Rescue effect of PKMYT1AR over-expression on miR-485-5p mimics-mediated phenotype was examined by tumor sphere formation assay (k), l is the quantification data for (k). Scale bar=50 μm. m-n Rescue effect of PKMYT1 over-expression on miR-485-5p mimics-mediated phenotype was examined by tumor sphere formation assay (m), (n) is the quantification data for (k). o-p Rescue effect of PKMYT1 over-expression on PKMYT1AR depletion-mediated phenotype was examined by tumor sphere formation assay (o), (p) is the quantification data for (o). * P < 0.05, ** P < 0.01, *** P < 0.001