Figure 7
From: Pro-inflammatory endothelial cell dysfunction is associated with intersectin-1s down-regulation
LPS-induces ITSN-1s deficiency without affecting the EC survival. Control and LPS-treated ECs lysates were analyzed by Western blot (A) and qPCR (B) for ITSN-1s protein and mRNA expression. The ITSN-1s mRNA levels, relative to the internal control, cyclophilin, were evaluated in three separate experiments. C. TUNEL staining of ECs treated with 1 μg/ml of LPS for 48 hours show no evidence for apoptotic activity (c2). A positive control - siRNA ITSN-1s - transfected ECs is shown in c1. D. Quantitative assessment was obtained by counting the number of TUNEL-positive and TUNEL-negative cells in 25 high-powered fields and calculating the percentage of apoptotic cells amongst the total cell population. No significant difference was noted in the percentage of apoptotic cells between LPS-treated and control ECs (0.89% vs. 0.98%, p = .45). E. Mitochondrial fractions and post-mitochondrial supernatants prepared for LPS-treated cells were analyzed for cyt c reactivity.