Journal: Frontiers in Cellular Neuroscience

Article Title: Enhancing autophagy mitigates LPS-induced neuroinflammation by inhibiting microglial M1 polarization and neuronophagocytosis

doi: 10.3389/fncel.2025.1546848

Figure Lengend Snippet: Effect of autophagy on the expression of NF-κB pathway proteins in mouse brain tissues as well as on the expression of LC3 in microglial cells. The 3-MA, RAPA, and LPS were stereotactically injected into the mouse brain. After 24 h, the frontal and temporal lobe tissues near the injection site of the lateral ventricle, as well as hippocampal tissues, were collected for Western blot analysis. (A) The expression levels of LC3II/I, Beclin-1, p-NF-κB, p-IκB-α, NF-κB, and IκB-α were detected by Western blot. (B) Immunofluorescence was used to evaluate LC3 expression in microglial cells. Scale bar: 10 μm. All data are presented as the mean ± standard deviation; n = 3; * p < 0.05; ** p < 0.01 vs. control. # p < 0.05, ## p < 0.01.

Article Snippet: The membrane was blocked with 5% skimmed milk for 1 h, followed by overnight incubation with primary antibodies, including rabbit anti-mouse LC3 antibody (Proteintech), rabbit anti-mouse Beclin-1 antibody (Affinity, United States), rabbit anti-rat inhibitor of NF-κB (IκB)-α antibody (Merck, NJ, United States), rabbit anti-rat NF-κB antibody (Merck), rabbit anti-rat p-IκB-α antibody (Merck), rabbit anti-rat p-NF-κB antibody (Merck), CD86 (Wuhan USCN Sciences Co., Ltd., China), CD206 (Wuhan USCN Sciences), and tubulin (Hyclone) at 4°C.

Techniques: Expressing, Injection, Western Blot, Immunofluorescence, Standard Deviation, Control