Fig. 2: Effect of PCK2 silencing on EC morphology and membrane integrity.

a Quantification of cell area in control and PCK2^KD1 ECs in 5.5 versus 0 mM glucose (n = 5). b Representative immunofluorescence images of F-actin (phalloidin staining; white) in control and PCK2^KD1 ECs in 5.5 versus 0 mM glucose (n = 5). c qRT-PCR analysis of tight junction protein 1 (TJP1) and claudin-5 (CLDN5) mRNA levels in control and PCK2^KD1 ECs in 5.5 versus 0 mM glucose (n = 3). d Representative immunofluorescence images of VE-cadherin staining and quantification of VE-cadherin⁺ (dis)continuous junction length (graph right to the images) in control and PCK2^KD1 ECs in 5.5 versus 0 mM glucose (n = 3). White arrowheads show VE-cadherin⁺ discontinuous junctions. e, f Quantification of reticular structure area (e) and gap size index (f) on VE-cadherin immunostained control and PCK2^KD1 ECs in 5.5 versus 0 mM glucose (n = 3). g Trans-endothelial electrical resistance (TEER) measurements in proliferation-blocked (MitoC-treated) confluent control and PCK2^KD1 EC monolayers in 5.5 versus 0 mM glucose (glc) (n = 5). Asterisks and hashtags in (g) denote statistically significant differences between KD and control respectively at 5.5 mM and at 0 mM glucose. Data are mean ± s.e.m. Statistics: ANOVA (a, c–f) two-tailed t-test with Welch’s correction (g); * or ^#P < 0.05; ** or ^##P < 0.01; *** or ^###P < 0.001; **** or ^####P < 0.0001; ns not significant. Scale bars in (b, d) are 10 µm; AU, arbitrary units.