Fig. 2

DMHCA treatment significantly attenuate RIR damage. a HE staining and quantitative analysis of IPL thickness in retina tissue harvested 3 days post-RIR injury; n = 6–10. Scale bar = 50 μm. Sham, control C57BL/6J with sham operation; RIR, retinal ischaemia/reperfusion injury model mice; DMHCA, DMHCA treatment with RIR mice. All in vivo experiments shared this grouping system. b Representative immunofluorescence images of anti-RBPMS (red) and anti-TUJ1 (green) labelled RGCs on a flat mount retina. From Sham, RIR, and DMHCA-treated mice at 3 days post-RIR. Quantitative analysis of numbers of RGCs shows a neuroprotection effect of DMHCA; n = 6. Scale bar = 50 μm. c Representative immunofluorescence images of RGCs in the retinal frozen section. Consistent with images of flat mount retina; n > 3. Scale bar = 50 μm. d TUNEL staining (red) labelled level of apoptosis in retina, showing an anti-apoptosis effect of DMHCA in vivo; n = 3. Scale bar = 50 µm. e Western blot analysis of the indicated proteins, Cleaved-caspase3, protein levels were normalized to GAPDH levels. f Transcriptional levels of RGC-specific indices, including Brn3a, Brn3c and RBPMS, and programmed cell death-related mRNA, such as caspase 8 and GSDMD, in retinas were detected using qRT-PCR; n > 3