Figure 2.
Silencing of Lnc-CRAT40 inhibits CRC proliferation, migration and invasion in vitro. A. Schematic of CRISPR/Cas9-mediated knockout of lnc-CRAT40 in HCT15 cells using paired sgRNAs targeting sequences flanking the transcriptional locus, resulting in a 1.1 kb deletion. B. Identification of knockout clones by genomic PCR and confirmation of homozygous deletion by Sanger sequencing; off-target regions served as negative controls. C. qRT-PCR analysis of lnc-CRAT40 expression in SW480 and HCT15 cells after CRAT40 silencing or control transfection (left), and in control versus CRAT40 knockout HCT15 clones (right). D. Cell proliferation assessed by CCK-8 assay following lnc-CRAT40 knockdown or knockout. E, F. Wound healing and Transwell assays evaluating migration and invasion in knockdown or knockout cells compared with controls; quantification shown on right. Data represent mean ± SD from three independent experiments. NC, negative control; KO, knockout. *P<0.1, **P<0.01, ***P<0.001, ****p<0.0001.
