Fig 2: ASC combined with bpV(pic) promotes autophagy and attenuates apoptosis in rat acute SCI.Notes: (A) Representative immunofluorescence images of NeuN (green) and LC3B (red) staining in spinal cords from SCI rats from the different groups. The fields in the frame were enlarged on the right side. Scale bar =50 µm. (B) Quantification of the number of NeuN (green)-positive cells in (A), n=5. (C) Representative immunofluorescence images of Bcl2 (green) and Bax (red) staining in spinal cords from SCI rats from the different groups. Scale bar =50 µm. (D) Quantification of the Bax/Bcl2 ratio in (C), n=5. (E) Immunoblot analysis of p62, LC3B, Beclin1, Bax, Bcl2, and GAPDH levels in spinal cords from SCI rats with different treatments. n=3 independent experiments.*In comparison with control group, P<0.05. The red * in (B) means P<0.05 between bpV(pic) and ASC combined with bpV(pic) group. #In comparison with ASC group, P<0.05. ns = not significant. All data are shown as mean ± SD.Abbreviations: ASC, acellular spinal cord; bpV(pic), bisperoxovanadium; LC3B, light chain 3B; SCI, spinal cord injury.

Fig 3: Treatment with bpV(pic) or ASC scaffolds combined with bpV(pic) promotes autophagy and attenuates apoptosis in cultured rat neurons.Notes: (A) Top, representative images of cocultured RNSCs with vehicle (Ctrl), ASC only, bpV(pic), or ASC combined with bpV(pic). Below, immunofluorescence analysis of LC3B expression (red) in RNSCs from different groups. Scale bar =50 µm. (B) RNSCs were cocultured with different materials for 24 hours, and then cell survival was analyzed by CCK-8 assay. n=3 independent experiments. (C) Quantification of the number of LC3B (red)-positive cells in (A), n=5. (D) Immunoblot analysis of p-PTEN, p62, Beclin1, LC3B, and GAPDH levels in RNSCs with different treatments. n=3 independent experiments. (E) Immunoblot analysis of cleaved-PARP, caspase 3, Bax, Bcl2, and GAPDH levels in RNSCs with different treatments. n=3 independent experiments. *In comparison with control group, P<0.05. The red * in (C) means P<0.05 between bpV(pic) and ASC combined with bpV(pic) groups. #In comparison with ASC group, P<0.05. ns = not significant. All data are shown as the mean ± SD.Abbreviations: ASC, acellular spinal cord; bpV(pic), bisperoxovanadium; CCK-8, cell counting kit-8; LC3B, light chain 3B; RNSCs, rat neuron stem cells; SCI, spinal cord injury.