Fig. 1: miR-200 FKO induces significant morphological alteration in the human GC cell line, AGS.
From: Complete loss of miR-200 family induces EMT associated cellular senescence in gastric cancer
A The multiplex CRISPR/Cas9 in a single lentiviral vector containing four sgRNAs driven by individual promoters. B Four sgRNAs indicated by scissors targeting distinct genomic loci of human miR-200 family host genes. C qPCR analysis of hsa-mir-200c-3p and miR-429 expression levels as a proxy in 39 single cells derived clonal population of AGS. D Further examination of the other miR-200s by qPCR in selected single-cell clones from C with miR-21 as a control. E Representative images of bright field (BF) and crystal violet staining illustrates morphological differences of A-NTC cells, the miR-200 FKO clone A-31, and A-26 with residual miR-200s expression from AGS. Scale bar, 100 μM. F F-actin (red)/DAPI (blue) and ZO-1 (green) staining of these three AGS clones. Scale bar, 20 μM. G Quantification of F-actin and Zo-1 fluorescent intensity from F. H Flow cytometry analysis of cell size from indicated clones. Data in D, G, and H are presented from triplicate analyses as the mean ± SEM. *p < 0.05, and **p < 0.01.
