AQP1 ablation mimicks M2 functional differentiation induced in WT macrophages by IL4/IL13: A- CD206 redistribution into ruffles. (A) Confocal images.

<p><i>Aqp1</i>^<i>+/+</i> and <i>Aqp1</i>^-/- macrophages were maintained for 24h in serum-containing medium without supplements (to keep macrophages in undifferentiated M0 state; upper row), or with LPS (to orient towards M1 phenotype; middle row) or IL4/IL13 (to orient towards M2 phenotype; lower row). Cells were fixed/permeabilized and double-immunolabelled for CD206 (green) and F4/80 (red) by reference to nuclei (Topro-3; blue). Representative of 2 independent experiments. Insets of M0 (<i>Aqp1</i>^<i>-/-</i>; panel b) and M2 macrophages (<i>Aqp1</i>^<i>+/+</i> and <i>Aqp1</i>^-/-; panels e and f) show strong recruitment of CD206 (green) into ruffles, contrasting with preferential perinuclear localization in WT M0 and M1 macrophages (panels a and c). Scale bars, 5μm. <b>(B) Morphometry: redistribution of CD206 in ruffles.</b> From left to right, analysis was performed on 54, 35, 30, 31, 10 and 7 cells, pooled from 2 independent experiments. NS, not significantly different; ***, p<0.001.