Figure 4: Fractalkine is required for Norgestrel’s neuroprotective effects against rd10 microglia in vitro.
From: Fractalkine-CX3CR1 signaling is critical for progesterone-mediated neuroprotection in the retina
(A) Example images of fractalkine immunofluorescence (red) in 661 W cells treated with siRNA against fractalkine. Scale bar 30 μm. (B) Western blot showing decreased levels of fractalkine in 661 W cells treated with siRNA fract. vs scrambled. Total protein level is shown alongside. (C) Detection of fractalkine mRNA levels by RT-PCR in 661 W cells following treatment with scrambled or siRNA targeted to fractalkine. (D) Cell viability of 661 W cells following siRNA fract. treatment as assessed by the MTS assay (N = 10 technical replicates). (E) Quantification of TUNEL+ 661 W cells pre-treated with a combination of scrambled RNA, siRNA fract., 20 μM Norgestrel or vehicle (DMSO) and in co-culture with rd10 microglia (N = 8 mice for primary culture, n = 6 technical replicates). (F) Example images of TUNEL+ 661 W cells (green) pre-treated with a combination of scrambled RNA, siRNA fract., Norgestrel or vehicle and in co-culture with rd10 microglia. Scale bar 30 μm. Hoechst reveals cell nuclei. Results are presented as mean ± SEM (two-way ANOVA, *p < 0.05, **p < 0.01, ****p < 0.0001).
