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Fig. 5 | Journal of Neuroinflammation

Fig. 5

From: Lipopolysaccharide induces neuroinflammation in microglia by activating the MTOR pathway and downregulating Vps34 to inhibit autophagosome formation

Fig. 5

Rapamycin alleviates the LPS-induced neuroinflammation in N9 microglial cells. a Treatment of N9 microglial cells with rapamycin enhances autophagy and facilitates the colocalization of GFP-DFCP and WIPI2-RFP even after stimulation with 1 μg/mL LPS for 12 h. Microglial cells were transfected with GFP-DFCP1 and RFP-WIPI2 plasmids for 24 h and then treated with 100 nM rapamycin with or without 1 μg/mL LPS for 12 h. Images were obtained by confocal microscopy. Scale bar: 10 μm. b Pearson’s correlation coefficient for colocalization of DFCP1 with WIPI2 was calculated by Image J. N9 microglial cells were treated with vehicle, LPS (1 μg/mL), rapamycin (100 nM) + LPS, rapamycin + LPS + 3-MA (2.5 mM), or LPS + 3-MA for 6 h. The levels of mRNAs encoding the proinflammatory cytokines TNF-α (c), IL-6 (d), and iNOS (e) were analyzed by qRT-PCR. Data are presented as mean ± SEM. ***p < 0.001 vs control; ##p < 0.01, ###p < 0.001 vs LPS; $$p < 0.01, $$$p < 0.001 vs cotreatment with LPS and rapamycin. f Effects of different conditioned media on the viability of SH-SY5Y cells assessed by MTT. Conditioned media (CM) were obtained from N9 microglial cells treated with or without LPS and rapamycin as indicated. Data are presented as mean ± SEM. ###p < 0.001 vs CM-CTRL; ***p < 0.001 vs CM-LPS

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