Figure 4

Effect of LRRK2-G2019S on phagocytosis and cytokines secretion in stimulated iMGL. The phagocytosis of iMGL was assessed using fluorescent pHrodo beads and Incucyte S3 Live-Cell Analysis System. (a) Representative images of the pHrodo phagocytosis and fluorescence intensity after (0 h and 5 h timepoints) in vehicle and IFNγ stimulated iMGLs. The phagocytosis was analyzed by measuring green fluorescence intensity. Purple color represents the pHrodo mask in the analysis. (b, c) Representative iMGL phagocytosis curves measured by green fluorescence intensity at the basal level and in IFNγ stimulated in iMGL. n = 1 batch, with 5 technical replicates (wells), presented as mean ± SD. (d, e) The phagocytosis of iMGL measured from pHrodo intensity at 5 h timepoint after stimulation with IFNγ, LPS or a combination of LPS + IFNγ. The results were normalized to vehicle treated control line (f-ISO1 or m-CTRL = 100%) for each batch of iMGL. n = 5 independent batches, with 5 technical replicates (wells) in each batch. The secretion of cytokines from iMGL in response to stimulation with IFNγ, LPS or LPS + IFNγ was measured from medium samples measured with cytokine bead array. (f) Secretion of IL8, IL10 and TNFα in female iMGL and (g) male iMGL. The cytokines were analyzed from medium samples collected from 4 (f-PD1, f-ISO1) or 3 (m-PD, m-CTRL) independent batches, with 5 technical replicates (wells). Data are presented as mean ± SD. Two-way Anova with Sidak ´s multiple comparison test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.