Rapamycin-induced autophagy quantitatively and functionally improved BM ECs from PGF patients. (a) Representative images (left panel) of BM ECs. Typical ECs from PGF and GGF patients were characterised by double-positive staining (merge, yellow) with DiI-AcLDL (red) and FITC-UEA-I (green) at day 7 of culture (original magnification, 10 × , scale bars represent 50 μm). Quantification (right panel) of double-positive BM ECs (merge, yellow) stained with DiI-AcLDL (red) and FITC-UEA-I (green) after 6 h of treatment with rapamycin (original magnification, 10 × , scale bars represent 50 μm). The (b) cell count, (c) intercellular ROS levels and (d) apoptosis rate of BM ECs from PGF and GGF patients were analysed after 6 h of treatment with rapamycin. Wilcoxon's test for paired data was used to identify drug effects. All P-values <0.05 were considered significant and are provided in the figure. *P<0.05, **P<0.005, ***P<0.0005.