Autophagy is suppressed by LPS in microglia. (a) Analysis of autophagic flux in primary microglia after treatment with the indicated dosages of LPS for 24 h. Graph, quantification of MAP1LC3B-II after normalization to ACTB (n = 3). (b) Analysis of autophagic flux in BMDMs after treatment with the indicated dosages of LPS for 24 h. Graph, quantification of MAP1LC3-II after normalization to ACTB (n = 3). (c) Analysis of autophagic flux in BV-2 microglial cells after treatment with LPS (1 μg/mL) for 6 or 24 h. (d) Analysis of autophagic flux in Raw264.7 macrophages after treatment with LPS (1 μg/mL) for 6 and 24 h. (e) Quantification of ZFYVE1 puncta following LPS treatment in BV-2 cells (n > 30 cells per condition). (f) Quantification of ZFYVE1 puncta following LPS treatment in Raw264.7 cells (n > 40 cells per condition). (g) Quantification of GFP-MAP1LC3B puncta in BV-2 cells 24 h after LPS (1 μg/mL) treatment (n > 30 cells per condition). (h) Time course analysis of MAP1LC3B-II level in primary microglia. Cells were treated with LPS (1 μg/mL) for 1, 2, 6, 12, or 24 h. Graph, quantification of MAP1LC3B-II after normalization to ACTB (n = 4). In all experiments where BafA1 was used, BafA1 (20 nM) was added 2 h before sampling. Scale bars: 10 μm. All data are mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001 compared to the control (Con) unless indicated otherwise.