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. Author manuscript; available in PMC: 2020 Sep 1.
Published in final edited form as: J Diabetes Complications. 2019 May 29;33(9):668–674. doi: 10.1016/j.jdiacomp.2019.05.016

Figure 5. Act1-FADD dependent retinal endothelial cell death.

Figure 5.

A) Immunoblot analysis of FADD expression in protein lysates of IL-17A stimulated scrambled control (Act1 expressing (ctrl)) or siAct1 (Act1 gene silenced) hREC cultures; protein was collected 6h post-stimulation. β-actin was used as a loading control. B) FADD expression by RT-PCR verifying gene silencing (siFADD) in hREC cultures; ctrl samples were transfected with scrambled sequences. Representative images of cellular death in IL-17A stimulated control (FADD expressing) and siFADD (FADD knockdown) hREC (scale bars of all images = 450μm) examined by microscopy (C) and quantified by flow cytometry (D) using Propidium Iodide (PI). Quantification of percent positive cells per flow cytometry analysis (n=6) of active Caspase 8 (E), Caspase 3 (F), and Annexin V (G) in IL-17A stimulated control (grey) and IL-17A stimulated siFADD (black) hREC cultures 6h after incubation. *=p<0.01 per unpaired student’s t-test. Data are representative of two separate experiments with similar results.