Fig. 5
Lrrk2 −/− primary microglia cells reported increased levels of phosphorylated and nuclear NF-κB p50 subunit under unstimulated conditions. a Lrrk2 +/+ and Lrrk2 −/− microglia lysates were subjected to immunoblotting using NF-κB P-p50 and p50 total antibodies. b Quantification of P-p50 subunit is normalized for p50 total protein. Data are representative of three independent experiments (bars represent the mean ± SEM; unpaired t test; **p > 0.01). c Lrrk2 +/+ and Lrrk2 −/− brain lysates were subjected to immunoblotting using NF-κB P-p50 and p50 total antibodies. d Quantification of P-p50 subunit is normalized for p50 total protein. Data are representative of three independent experiments (bars represent the mean ± SEM; unpaired t test; *p > 0.05). e BV2 cell lysates treated with GSK or DMSO as control were subjected to immunoblotting using NF-κB P-p50 and p50 total antibodies. f Quantification of P-p50 subunit is normalized for p50 total protein. Data are representative of three independent experiments (bars represent the mean ± SEM; unpaired t test; *p > 0.05). g Representative images of unstimulated Lrrk2 +/+ and Lrrk2 −/− microglia cells stained for NF-κB p50 (red) and nuclei with Hoechst (blue). Scale bar 10 μm. h Quantification of nuclear NF-κB p50 shown as mean of fluorescence intensity from four independent experiments (~90 cells per experiment per genotype). Nuclear NF-κB p50 was calculated as nuclear fluorescence intensity divided by the nuclear area (μm2) (bars represent the mean ± SEM; unpaired t test; *p < 0.05). i Cumulative frequency distributions of nuclear p50 in Lrrk2 +/+ and Lrrk2 −/− microglia cells (n (Lrrk2+/+) = 352 and n (Lrrk2−/−) = 377; Kolmogorov-Smirnov test, p = 0.004). j BV2 cell lysates treated with forskolin or PKI were subjected to immunoblotting using P-p50 and p50 total antibodies. k Quantification of P-p50 is normalized for p50 total protein. Data are representative of five independent experiments (bars represent the mean ± SEM; unpaired t test; *p > 0.05, ***p > 0.001)