FIG 3.

Rab5a knockout inhibits CdtB-induced cytotoxicity and apoptosis in human colonic epithelial cells. Stable primary human colonic epithelial cells, with the lenti-CRISPR/Cas9-GFP Rab5a-KO construct (“Rab5a-KO” cells) or the empty vector (“CRISPR-C”) were treated with Cdt holotoxin (2 μg/ml) for the applied time; the expression of the listed genes was tested by qPCR (A) and Western blotting (B); cell viability and cell death were tested by an MTT assay (C) and an LDH medium release assay (D), respectively; cell apoptosis was quantitatively analyzed by the annexin V-FACS assay (E). *, P < 0.05 versus Ctrl treatment of CRISPR-C cells. ^#, P < 0.05 versus CdtB treatment of CRISPR-C cells. The experiments reflected in this figure were repeated three times, and similar results were obtained.