Fig 2: TNFa and CHX trigger an apoptotic response in cFLIP knockdown FHL124 cells by activating caspase-8, caspase-9, and caspase-3.A Activated caspase-8 was detected in cFLIP KD3 cFLIP knockdown FHL124 cells from 5 h to 24 h TNFa and CHX treatment. Similar results were seen in activated caspase-9 and caspase-3. PARP-1 cleavage was detected at 7 h and 24 h time point after the stimulation. B–E The semi-quantitative densitometric measurement. One-way ANOVA was used to compare between groups, and only p < 0.05 is considered significant. *<0.05, **<0.01, ***<0.001, ****<0.0001.

Fig 3: A model of cFLIP-mediated LEC’s survival.A proposed model is highlighting cFLIP regulation of lens epithelial cells’ survival and anti-apoptotic function.

Fig 4: Synergistic effects of trichostatin A (TSA) and cisplatin on Cellular Fas-associated death domain-like interleukin-1 ß converting enzyme inhibitory protein (cFLIP) as detected by Western blotting. 1: control; 2: TSA 250 nmol/L; 3: cisplatin 10µg/mL; 4: TSA 250 nmol/L+ cisplatin 10µg/mL. * p < 0.05,**p < 0.01 versus control group.

Fig 5: TNFa and CHX trigger an apoptotic response in cFlip knockdown 17EM15 cells and ex vivo cultured mouse lens capsular bag by activating caspase-8, caspase-9, and caspase-3.A cFlip knocking down in 17EM15 mouse LECs determined by immunoblot-assay. B The semi-quantitative densitometric measurement of 17EM15 cFlip knocking down from immunoblot-assay. C cFlip knocking down in ex vivo cultured mouse lens capsular bags. D The semi-quantitative densitometric measurement of cFlip knocking down mouse lens capsular bags from immunoblot-assay. E Pro-caspase-8, cleaved caspase-8, pro-caspase-3, and cleaved caspase-3 in cFlip knockdown and scrambled shRNA control (SC) 17EM15 cells with and without 30 ng/ml TNFa and 10µg/ml CHX stimulation for 7 h. F Pro-caspase-8, cleaved caspase-8, pro-caspase-3, and cleaved caspase-3 in cFlip knockdown and scrambled shRNA control (SC) ex vivo cultured mouse lens capsular bags with and without 60 ng/ml TNFa and 10 µg/ml CHX treatment for 24 h. The cFlip KD 17EM15 treated cell lysate was used as a positive control. Only 1/10th amount of protein relative to capsular bag lysate was loaded. One-way ANOVA was used to compare between groups, and only p < 0.05 is considered significant. *<0.05, **<0.01, ***<0.001, ****<0.0001.