Journal: Nature Communications
Article Title: CRISPR-CasRx-mediated disruption of Aqp1/Adrb2/Rock1/Rock2 genes reduces intraocular pressure and retinal ganglion cell damage in mice
doi: 10.1038/s41467-024-50050-4
Figure Lengend Snippet: a , b IOP changed during day and night in each group after monocular magnetic beads modeling or DEX modeling and injection of shH10 Y25 virus. WT, n = 20; Beads+saline n = 10; Beads+LacZ n = 8; Beads+Y25 n = 20. DEX+saline n = 20, DEX+LacZ n = 18, DEX + Y25 n = 40. c – f The retinas of microbeads model mice or Dex-induced model mice treated with shH10 Y25 virus were taken 10 weeks after modeling, and the RGC counting results of Rbpms+ were compared among all groups. Each retina takes 6–12 visual fields, including center, mid-peripheral and peripheral positions, Scale bars, 100 μm. c and e RGC count results of three retinal regions in each microbeads model group. In peripheral, Y25 compared with LacZ, P < 0.0001. WT n = 8, Beads + saline n = 6, Beads + LacZ n = 5, Beads+Y25 n = 8. d and f RGC count results of three retinal rings in each Dex-induced model group. In peripheral, Y25 compared with LacZ, P < 0.0001. WT n = 6, DEX+saline n = 10, DEX+LacZ n = 6, DEX + Y25 n = 11. g , h Diurnal and nocturnal IOP curves of mice in each group monocular microbeads modeling or DEX modeling and injection of shH10 Y26 (AAV-EFs-CasRx/U6- Aqp1 - Adrb2 ) virus. WT group n = 18, Beads+saline group n = 8, Beads+LacZ group n = 7, Beads+Y26 group n = 19. WT n = 20, DEX+saline n = 18, DEX+LacZ n = 20, DEX + Y25 n = 34. i RGC counting results of center, mid-peripheral and peripheral retinal rings were obtained in microbeads model mice treated with shH10 Y26 virus. In peripheral, Y26 compared with LacZ, P = 0.0001. WT n = 6, Beads+saline n = 5, Beads+LacZ n = 5, Beads+Y26 n = 6. j RGC counting results of three retinal rings were obtained in Dex-induced model mice treated with shH10 Y26 virus. In peripheral, Y26 compared with LacZ, P = 0.0006. WT n = 5, DEX+saline n = 4, DEX+LacZ n = 5, DEX + Y26 n = 7. All values were expressed as mean ± SEM. The differences of IOP between groups were tested by one-way ANOVA. Other data were statistically analyzed by two-tailed, unpaired T -test. Compared with WT, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; Compared with LacZ, # P < 0.05, # # # # # P < 0.01, # # # # P < 0.001.
Article Snippet: After washing with PBS for 5 min, retinas were incubated with RBPMS rabbit polyclonal antibody (1:100; Cat#15187-1-AP; Proteintech, USA) overnight at 4 °C and washed 3 × for 5 min each with PBS before 2 h of incubation with secondary antibodies at room temperature.
Techniques: Magnetic Beads, Injection, Virus, Saline, Two Tailed Test