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. 2022 Apr 27;11(9):1468. doi: 10.3390/cells11091468

Figure 4.

Figure 4

ANRIL/p15 regulates cell senescence in SIG. (A) Biological pathways participating in SIG were analyzed using Panther enrichment analyses. Disturbed genes in SIG TM samples compared to controls were enriched in the deposit of ECM and cellular senescence. n = 3 per group. (B) TM samples from control and SIG mice were prepared as single cell suspensions and labeled with PI for cell cycle analysis. In SIG TM samples, there was a huge increase of cells arrested in G1 stage and decrease in other stages. n = 5 per group. (C) In cultured mouse TM cells, DEX treatment induced G1 arrest by blocking more cells in the G1 phase compared to vehicle treated cells. ANRIL depletion also induced G1 arrest as DEX. Depletion of p15 prevented DEX induced G1 arrest. n = 6 per group. (D) In cultured mouse TM cells, the expression of p15 protein and Cyclin D3 protein was measured in nuclear protein fractions, while Histone H3 served as internal controls. The phosphorylated Rb protein was checked in cytoplasm fraction with total Rb protein as control. n = 6 per group. (E) Quantification of (D). Notes: CON = control; SIG = steroid-induced glaucoma; ECM = extracellular matrix; Veh = vehicle; siANRIL = ANRIL knock down; DEX = dexamethasone; sip15 = p15 knock down; ** p < 0.01 compared with vehicle; # p < 0.05 compared with scramble RNA; ## p < 0.01 compared with scramble RNA; NS = no significant difference.