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. 2020 Jan 31;10:1597. doi: 10.1038/s41598-020-58482-w

Figure 3.

Figure 3

Immunoblot analysis of the protein expression of Lc3b-II, Lc3b-I, p62 and Beclin1 in PC12 cells treated with the indicated PHD inhibitors (100 µM) and rapamycin (1 & 10 µM) for 24 hours in normoxia. (A) Representative immunoblots of Lc3b-II, Lc3b-I, p62 and Beclin1 were shown alongside β-actin; (B) Normalised Lc3b-II/Lc3b-I ratio measured after 24 hours exposure to the indicated PHD inhibitors and rapamycin in normoxia (n = 3). Significant increase in the Lc3b-II/Lc3b-I ratio was seen with rapamycin (1 & 10 µM) and 100 µM of DMOG, FG2216, FG4592, GSK1278863 and Bay85-3934. GSK1278863 and Bay85-3934 had a similar effect on the Lc3b-II/Lc3b-I ratio as Rapamycin; (C) Expression of p62 measured after 24 hours exposure to the indicated PHD inhibitors and rapamycin in normoxia (n = 3). Significant reduction in p62 expression in comparison to vehicle (1% DMSO)-treated cells was seen with rapamycin (1 & 10 µM) and 100 µM of DMOG, FG2216, FG4592, GSK1278863 and Bay85-3934 treated cells; (D) Expression of Beclin1 measured after 24 hours exposure to the indicated PHD inhibitors and rapamycin in normoxia (n = 3). Significant upregulation of Beclin1 expression in comparison to vehicle (1% DMSO)-treated cells was seen in 100 µM of DMOG, FG2216, FG4592, GSK1278863 and Bay85-3934 treated cells, but was not in the Rapamycin treated cells. Data were expressed as mean ± S.D. *Indicated P < 0.05 against 1% DMSO treatment (Two-way ANOVA, Tukey’s post-hoc analysis).