Skip to main content
. 2019 Jan 11;14(1):e0210617. doi: 10.1371/journal.pone.0210617

Fig 2. The effects of mTOR inhibitor rapamycin on autophagy in GD and FD.

Fig 2

(A-C) PBMCs from healthy controls (A), GD (B), and FD (C) were treated with the 10 nM rapamycin (RAP) alone or in combination with 20 μM 3MA for 3h. Then, PBMCs were stained with Cyto-ID autophagy detection kit. Samples were measured in triplicates, control group n = 5, GD1 n = 11, GD3 n = 3, FD n = 5. All data were expressed as S.E.M. and * p<0.05 vs control. (D) Representative western blot showing Beclin1 and LAMP1 protein expression in GD (N370S/L444P and L444P/L444P mutations), FD, and control samples after 3h treatment with 10 nM rapamycin. (E) Quantification of relative levels of Beclin1 and LAMP1 normalized to actin from separate experiments in which the control were untreated PBMCs from the same group. n = 5–7 samples analysed each patient group. Membranes were re-probed with actin or total protein for normalization.* p<0.05 vs untreated samples. (F) Q-PCR was performed to determine relative DDIT3 and HSPA5 mRNA expression levels in GD and FD samples. The RNA level was measured in triplicates, n = 5 or 6 samples analysed each group. All data are expressed as S.E.M. and * p<0.05 vs untreated samples.