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Fig. 8 | Journal of Neuroinflammation

Fig. 8

From: Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTPσ receptors promotes a beneficial inflammatory response following spinal cord injury

Fig. 8

CSPGs modulate microglia phagocytosis, migration, and nitrite production which is partially mediated through LAR and PTPσ signaling and Rho activation. a, b Microglia phagocytosis was assessed. Success of phagocytosis was verified by intracellular detection of green fluorescent beads in microglia (Iba-1+) through Z-stack imaging. M1 microglia (TNFα + IFNγ treated) showed a reduced ability for phagocytosis. CSPGs reduced phagocytosis in microglia, which was attenuated and even promoted with ILP/ISP, inhibition of ROCK by Y-27632, or ChABC treatment but not by TAT or IMP control peptides. c Representative images of phagocytosis by M0, M1, and M2 microglia are depicted for PDL, CSPGs, and CSPGs + ILP/ISP conditions. d Nitrite production was exacerbated and significantly increased in M1 microglia when exposed to CSPG. This effect was not blocked by ILP/ISP, Y-27632, IMP, or TAT treatment but was blocked by ChABC degradation of CSPGs. IL-10 (e) and IL-1β (f) release was assessed in microglia 2 days after plating onto PDL or PDL + CSPGs substrate. CSPGs reduced IL-10 expression in M2 microglia while had no significant effect on IL-1β release. g CSPGs also significantly limited microglia migration which was overcome by ILP/ISP, Y-27632, and ChABC treatment. h RhoA activity was assessed by G-LISA in microglia demonstrating a significant increase in Rho activity when microglia were exposed to CSPGs substrate. ILP and ISP treatment significantly decreased Rho activity. The data show the mean ± SEM, *p < 0.05, one-way ANOVA, N = 3–5/group. N = 3–5 independent experiments. The data show the mean ± SEM, *p < 0.05, one-way ANOVA

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