Figure 1 | Scientific Reports

Figure 1

From: Hypoxia-inducible factor (HIF) prolyl hydroxylase inhibitors induce autophagy and have a protective effect in an in-vitro ischaemia model

Figure 1

PC12 cells were cultured in normoxia (21% O2) for 24 hours with the PHD inhibitors (FG4592, FG2216, GSK1278863, Bay85-3934; concentration of 1, 10, 50, 100 µM) and DMOG (1, 10, 50, 100, 250, 500 µM, 1 and 2 mM). (A) MTT assays (n = 3) reveals reduction in mitochondrial activity (MTT release) in cells subjected to 100 µM of FG4592 and 50 & 100 µM of Bay85-3934 in comparison to 1% DMSO. The other inhibitors did not significantly affect mitochondrial activity. A significant reduction in MTT release was seen by 250, 500 µM, 1 and 2 mM of DMOG in comparison to 1% DMSO; (B). LDH assays (n = 3) revealed no significant changes in structural integrity (% LDH release) with FG4592, FG2216, GSK1278863, Bay85-3934 at all concentrations. DMOG increased % LDH release and resulted in significant cytotoxicity at concentration of 500 µM, 1 and 2 mM; (C). Trypan blue exclusion assay (n = 3) revealed no significant changes in % of live (trypan blue unstained cells) with FG4592, FG2216, GSK1278863, Bay85-3934 at all concentrations. DMOG significantly decreased % of live cells at concentration of 500 µM, 1 and 2 mM. Data were expressed as mean ± S.D. *Indicates P < 0.05 against 1% DMSO treated PC12 cells (Two-way ANOVA, Tukey’s post-hoc analysis).

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