Fig. 5
Blocking CCR2 with INCB3344 inhibits calcium-mobilization induced by CCL2 in DRG neurons from CFA rats. DRG neuronal cells were loaded with 2 μm Fura-2-AM and monitored for relative fluorescence changes. a Representative images of calcium imaging recording of DRG cell culture from rats, 3 days post-CFA. Digital images (×40) were taken immediately before addition of CCL2 (left), after the addition of 50 nM CCL2 (middle), and after treatment with 1 μM of the calcium ionophore ionomycin, used as a positive control (right). Graded color scales indicate Fura-2-AM emission ratios (340/380 nm), with purple representing low [Ca2+]i and yellow/red representing high [Ca2+]i. Pre-incubation of 100 nM INCB3344 (30 min) blocks the rise of calcium induced by CCL2 application at day 3 post-CFA (b) and partially inhibits calcium-released at day 10 post-CFA (c). Quantification of the calcium mobilization induced by CCL2 in the presence or absence of INCB3344 (d). Percentage of cells responding to CCL2 in the presence or absence of INCB3344 (e). Cells were considered responsive when we observed at least 30% of increase in fluorescence intensity ratio, compared to baseline. Data shown correspond to mean ± SEM. # P < 0.05, *** P < 0.001 #### P < 0.0001; one-way ANOVA followed by Bonferroni post-test