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Fig. 1 | Journal of Neuroinflammation

Fig. 1

From: Loss of Par1b/MARK2 primes microglia during brain development and enhances their sensitivity to injury

Fig. 1

Knockdown of Par1b significantly activates primary microglia. a Primary microglia were transfected with shRNA targeting either luciferase or Par1b shRNA. Seventy-two hours post-transfection, cells were serum starved, stimulated with 50 μM ATP, fixed, and immunostained with Rhodamine-Phalloidin (red) and DAPI (blue). Scale bar = 50 μm. b Quantification of microglial circularity (4π × area/perimeter2) under conditions described in a. n = 472 cells for Lucif-shRNA Ctrl, 542 cells for Lucif-shRNA ATP, 522 cells for Par1b-shRNA Ctrl, and 483 cells for Par1b-shRNA ATP, from three independent experiments, ***p < 0.001 by two-way ANOVA. c Primary microglia were transfected with different shRNAs as in a. Seventy-two hours post-transfection, cells were serum starved, incubated for 2 h with UV damaged neurons, fixed, and immunostained with TuJ1 (red) and DAPI (blue). Scale bar = 5 μm. d Orthogonal view shows that the TuJ1+ particles are within the microglia. Scale bar = 5 μm. e Quantification of the number of TuJ1+ particles per cell. n = 37 cells for Lucif-shRNA and 34 cells for Par1b-shRNA from two independent experiments, ***p < 0.001 by one-way ANOVA

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