Figure 4.

Hypoxia and glucose deprivation synergise to inhibit ROCK signalling in endothelial cells. Confluent cultures were switched to normal DMEM or DMEM without glucose (Glc), supplemented with 5% dialysed FCS and then immediately exposed to 21%, 1% or 0.1% O₂. After 5 h, cells were treated with CA4P (1 μM) for a further 15 min. (a) Cell extracts were analysed for the indicated proteins. (b) pMLC was normalised to actin and expressed as fold increase over control cells maintained in 21% O₂. (c) Cells were stained for F-actin with Texas Red phalloidin (red) and β−tubulin (green). Blebbing cells are indicated with white arrows. Results are from 3 independent experiments. Data were analysed by one-way Anova followed by a Tukey post-test. Values are means ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).