FIG 5.

Rab5a is required for CdtB-induced proinflammatory responses in THP-1 human macrophages. Stable THP-1 human macrophages, with Rab5a shRNA (“S2”) or the lenti-CRISPR/Cas9 Rab5a knockout construct (Rab5a-KO cells), as well as the parental control THP-1 macrophages (“C”), were treated with Cdt holotoxin (2 μg/ml) for the applied time; the expression of the listed genes was tested by qPCR (A, D, and E) and Western blotting (B and I); cell viability was tested by an MTT assay (C); the production of listed cytokines in the medium supernatants was tested by ELISA (F and G); p65 DNA-binding activity was tested as well (H). *, P < 0.05 versus Ctrl treatment of C cells. ^#, P < 0.05 versus CdtB treatment of C cells. The experiments reflected in this figure were repeated three times, and similar results were obtained.