Heat shock pretreatment improves stem cell repair following ischemia-reperfusion injury via autophagy

Figure 2 Autophagy was determined by acidic vesicular organelle-positive mesenchymal stem cells (labeled in the circle) using a fluorescent dye (AO) and flow cytometry. A: Different periods of heat shock pretreatment (HSP) ranging from 1 to 3 h led to a time-dependent increase in autophagy in mesenchymal stem cells (MSCs) exposed to H₂O₂, which peaked in the HSP_3h group; B: 3-MA attenuated, whereas rapamycin failed to further increase HSP-induced autophagy; C: SB203580 significantly suppressed HSP-induced autophagy in MSCs exposed to H₂O₂; D: Western blot showed a significant time-dependent increase in expression of the autophagic marker LC3B-II and beclin 1 in MSCs. The data represent the results of three separate experiments. ^aP < 0.05, ^bP < 0.01 compared with the normal control group; ^cP < 0.05, ^dP < 0.01 compared with the HSP_2h control group.