Fig. 6

PDIA3 contributes to cell death from oxygen–glucose deprivation. a A 24 h period of OGD caused a loss of live (yellow arrows) and appearance of apoptotic (white arrowheads) CMECs compared to normoxic control conditions, as revealed by blue nuclear Hoechst staining. b Cell counts from 3 fields/well show that the number of live PDIA3-/- cells was four times higher than WT cells after OGD (p < 0.001, n = 3, Student’s t-test). c Pharmacological inhibition of STAT3 with stattic reduced the protective effect of PDIA3-/- against OGD in CMECs (Student’s t-test, n = 3, p < 0.05). d Western blotting shows that PDIA3-/- CMECs had more phosphorylated STAT3 S727, as measured by densitometry (e) as well as increased pFAK Y397 which was completely absent in control conditions or WT CMECs undergoing OGD. STAT3 Y705 phosphorylation was lost after OGD in both PDIA3-/- and WT CMECs, indicating Y705 plays no part in PDIA3-/- cell survival. Beta actin was used as a loading control to account for differences in cell survival and STAT3 and actin blots were run on the same gel. f PDIA3 expression was not different in WT control (Normoxia) relative to WT OGD CMECs as measured by densitometry (g)