p mtor s2448 cell signaling technology  (Cell Signaling Technology Inc)

Journal: Scientific reports

Article Title: Withaferin a modulation of microglia autophagy mitigates neuroinflammation and enhances cognitive function in POCD.

doi: 10.1038/s41598-024-75284-6

Figure Lengend Snippet: Fig. 5. WA promoted microglia autophagy and accelerated the degradation of LDs. Microglia were pre- treated with MG132 (10 µM) or 3-MA (5 mM) for one hour, followed by WA protection for one hour, and subsequently stimulated with LPS for 12 h. (A) BODIPY fluorescence probe was used to label LDs molecules in primary microglia (scale bar = 5 μm). (B) Average fluorescence intensity of labeled LDs was measured. (Four independent replicate experiments were conducted). (C) LDs accumulation was assessed in primary microglia through Oil Red staining (scale bar = 5 μm). (D Average area covered by labeled LDs on cells was determined (Four independent replicate experiments were conducted). (E) Primary microglia were labeled using a dual-fluorescence virus (GFP-RFP-MAP1LC3B) to visualize autophagosomes (scale bar = 5 μm). (F) Immunoblotting was performed to detect SQSTM1 and MAP1LC3B expression levels. Band intensities of SQSTM1 and MAP1LC3B in Western blots were quantified and presented in (G) and (H), respectively (Three independent replicate experiments were performed). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the corresponding group, as determined by the one-way ANOVA .

Article Snippet: For primary antibody incubation, the membrane was subjected to an overnight incubation at 4 °C (MAP1LC3B: Cell Signaling Technology, 2775, 1:1000; SQSTM1: Santa, sc-48402, 1:500; GAPDH antibody: Cell Signaling Technology, 5174, 1:3000).

Techniques: Fluorescence, Labeling, Staining, Virus, Western Blot, Expressing

Journal: Scientific reports

Article Title: Withaferin a modulation of microglia autophagy mitigates neuroinflammation and enhances cognitive function in POCD.

doi: 10.1038/s41598-024-75284-6

Figure Lengend Snippet: Fig. 6. WA improved the autophagy level in mice with POCD. (A) Co-labeling of microglia in the hippocampal region of mice with MAP1LC3B was performed (scale bar = 10–5 μm), analyzed by Imaris software and the quantified data were presented in (B) (N = 3). (C) Co-labeling of microglia in the hippocampal region of mice with SQSTM1 was carried out (scale bar = 40–20 μm)). The relative fluorescence intensity of SQSTM1 was shown in (D), and the number of co-labeled cells was displayed in (E) (N = 3).*P < 0.05, **P < 0.01, ***P < 0.001 compared with the corresponding group, as determined by the one- way ANOVA .

Article Snippet: For primary antibody incubation, the membrane was subjected to an overnight incubation at 4 °C (MAP1LC3B: Cell Signaling Technology, 2775, 1:1000; SQSTM1: Santa, sc-48402, 1:500; GAPDH antibody: Cell Signaling Technology, 5174, 1:3000).

Techniques: Labeling, Software, Fluorescence