Figure 4. Migration of MLS-9 microglial cells is increased by IL4 and IL10, and involves TRPM7, not KCa2.3.

A. Representative fluorescence micrographs show MLS-9 cells (rat microglial cell line), unstimulated or 24 hr after stimulation with LPS (100 ng/ml), IL4 (20 ng/ml) or IL10 (20 ng/ml). Cells were stained for F-actin with phalloidin (green) and the nuclear marker, DAPI (blue). Most cells were bipolar with membrane ruffling at one end (examples shown by arrows). Scale bar, 50 µm. B. MLS-9 microglial cells in the upper well of Transwells were unstimulated, or stimulated (24 hr) with 20 or 100 ng/ml IL4, or with 20 or 100 ng/ml IL10. The number of cells that migrated to the underside of the filter was then counted and normalized to control (unstimulated) cells. The dashed line in both graphs indicates the level in control (unstimulated) cells. C. Cells were stimulated as in panel B, with or without a channel inhibitor: 100 nM apamin, 5 nM tamapin, or 7 µM NS8593. For each stimulus, the number of cells that had migrated was normalized to the level without a channel inhibitor. Data are expressed as mean ± SEM with the number of individual cultures indicated on each bar. In panel B, a two-way ANOVA with Bonferroni post-hoc analysis was used to determine significant differences between stimulation (*) and dose (†). In panel C, A one-way ANOVA with Tukey's post-hoc test was used to determine treatment effects. One symbol, p<0.05; two symbols, p<0.01; three symbols, p<0.001.