Fig. 7
The effect of CoCl2 on transepithelial electrical resistance and transepithelial flux of 14C–D-mannitol of HIF-1β knockdown CaCo-2 cell layers. a Western immunoblot data showing the levels of HIF-1β protein in whole cell lysates of 4 control CaCo-2 cell layers (lanes 5–8) vs 4 HIF-1β knockdown cell layers (lanes 1–4). b Three-day-post-seeding control cell layers and 3 HIF-1β knockdown CaCo-2 cell layers (also 3-day-post-seeding) on Millicell PCF polycarbonate filters were refed in control medium or medium containing 125 μM CoCl2 on the apical and basal-lateral sides, 48 h prior to electrical measurements. Data shown represent the percent of respective control resistance and are expressed as the mean ± SEM of 8 cell layers per condition. After electrical measurements, radiotracer flux studies with 0.1 mmol/L, 0.1 μCi/mL 14C–D mannitol were performed on the same CaCo-2 cell layers for which resistance measurements were performed. Data shown represent the percent of respective control mannitol flux rate and are expressed as the mean ± SEM of 8 cell layers per condition. **P < 0.01, ***P < 0.001 vs cobalt treated control knockdown. (Student’s t test, two-tailed)