Journal: Journal of Neuroinflammation

Article Title: Microglia TFEB activation attenuates Alzheimer’s disease pathology by enhancing autophagy-lysosomal function

doi: 10.1186/s12974-026-03728-z

Figure Lengend Snippet: Generation and validation of 5xTFEB mice with microglia-specific hTFEB overexpression. a Breeding strategy for generating 5xTFEB mice by crossing 5xFAD mice with TFEB-CA transgenic lines. b Experimental workflow for isolation of microglial and non-microglia cell population from 6-month-old WT, 5xFAD, and 5xTFEB mice using MACS, followed by RT-qPCR and western blot analyses. c RT-qPCR analysis showing that endogenous mouse Tfeb mRNA levels were not significantly altered, whereas human TFEB ( hTFEB ) transcripts were robustly induced specifically in microglia from 5xTFEB mice (WT: n = 3, 5xFAD: n = 4, 5xTFEB: n = 4). d Western blot analysis of MACS-isolated microglia and non-microglia from WT, 5xFAD, and 5xTFEB mice using the indicated antibodies. e Immunofluorescence analysis of cortical and hippocampal sections from 6-month-old 5xFAD and 5xTFEB mice stained for hTFEB (green), Iba1-positive microglia (red), and GFAP-positive astrocytes (magenta). f Immunofluorescence analysis of cortical and hippocampal sections from 6-month-old 5xFAD and 5xTFEB mice stained for hTFEB (green), Iba1-positive microglia (red), and NeuN-positive neurons (magenta). Nuclei were counterstained with DAPI (blue). Scale bar, 10 μm. Data are presented as mean ± SEM. Statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparisons test (c). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Membranes were blocked with 3% BSA in Tris-buffered saline with Tween (TBS-T) and incubated overnight at 4 °C with primary antibodies against TFEB (Bethyl Laboratories, Montgomery, TX, USA), GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA), Lamin-B1 (Abcam), p62/SQSTM1 (Progen, Heidelberg, Germany), LC3 (Novus Biologicals, Littleton, CO, USA), human-specific TFEB (Cell Signaling Technology), AIF-1/Iba1 (Novus Biologicals, Englewood, CO, USA), anti-β-amyloid (1–16; Biolegend), Beclin1 (Cell Signaling Technology), Lamp1 (Abcam), NLRP3 (Adipogen, San Diego, CA, USA), ASC (Cell Signaling Technology), cleaved-caspase1 (Cell Signaling Technology), and interleukin (IL)-1β (Santa Cruz Biotechnology).

Techniques: Biomarker Discovery, Over Expression, Transgenic Assay, Isolation, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: Microglia TFEB activation attenuates Alzheimer’s disease pathology by enhancing autophagy-lysosomal function

doi: 10.1186/s12974-026-03728-z

Figure Lengend Snippet: Microglia-specific TFEB overexpression reduces amyloid plaque burden in 5xFAD mice. a Representative images of amyloid deposition in the cortex (CX), hippocampus (HP), and entorhinal cortex (EC) of WT, 5xFAD, and 5xTFEB male mice. Fibrillar amyloid plaques were visualized by Thioflavin-S (Thio-S, green) staining, and total Aβ plaques were detected by 6E10 immunostaining (red). No plaques were observed in WT brains. Quantification revealed a significant reduction in both fibrillar and total plaque burden in 5xTFEB compared with 5xFAD mice ( n = 6 per group, scale bar, 25 μm). b Western blot analysis of cortical lysates probed with 6E10 antibody showing reduced Aβ monomer levels in 5xTFEB mice relative to 5xFAD mice ( n = 4 per group). c ELISA quantification of cortical Aβ 1−42 confirming significantly lower levels in 5xTFEB compared to 5xFAD mice ( n = 6 per group). Data are presented as mean ± SEM. Statistical analyses were performed using Student’s t-test (a-b). * p < 0.05, ** p < 0.01

Article Snippet: Membranes were blocked with 3% BSA in Tris-buffered saline with Tween (TBS-T) and incubated overnight at 4 °C with primary antibodies against TFEB (Bethyl Laboratories, Montgomery, TX, USA), GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA), Lamin-B1 (Abcam), p62/SQSTM1 (Progen, Heidelberg, Germany), LC3 (Novus Biologicals, Littleton, CO, USA), human-specific TFEB (Cell Signaling Technology), AIF-1/Iba1 (Novus Biologicals, Englewood, CO, USA), anti-β-amyloid (1–16; Biolegend), Beclin1 (Cell Signaling Technology), Lamp1 (Abcam), NLRP3 (Adipogen, San Diego, CA, USA), ASC (Cell Signaling Technology), cleaved-caspase1 (Cell Signaling Technology), and interleukin (IL)-1β (Santa Cruz Biotechnology).

Techniques: Over Expression, Staining, Immunostaining, Western Blot, Enzyme-linked Immunosorbent Assay