Fig. 3

TA inhibits Aβ(1–42)-induced activation of the NLRP3 inflammasome via autophagy induction. A BV-2 cells were pretreated with 5 μM of Aβ(1–42) for 12 h, followed by treatment with TA at the indicated concentrations for an additional 24 h. Protein expressions were detected using Western blot. Bar charts indicate the ratios of NLRP3/β-actin, ASC/β-actin, caspase-1 (p10)/pro-caspase-1, IL-1β (p17)/pro-IL-1β, IL-18 (p18)/pro-IL-18, and GSDMD (p30)/pro-GSDMD in BV-2 cells. B BV-2 cells were pretreated with 5 μM of Aβ(1–42) for 12 h, followed by the treatment of TA (10 μM) in the presence or absence of LY (10 μM) for an additional 24 h. Protein expressions were detected using the Western blot. The bar chart indicates the ratios of NLRP3/β-actin, ASC/β-actin, caspase-1 (p10)/pro-caspase-1, IL-1β (p17)/pro-IL-1β, IL-18 (p18)/pro-IL-18, and GSDMD (p30)/pro-GSDMD in BV-2 cells. C Representative images of BV-2 cells transfected with pEGFP-NLRP3, mCherry-ASC, or pEGFP-caspase-1 plasmid for 18–24 h and treated with Aβ(1–42) in the presence or absence of TA at the indicated concentrations for 24h, followed by DAPI staining were captured. Magnification, × 10; scale bar: 100 μm. The bar chart indicates the percentage of BV-2 cells with NLRP3, ASC, or caspase-1 signal. D BV-2 cells seeded on confocal petri dishes were transiently transfected with pmCherry-C1-ASC and GFP-LC3 plasmid for 24 h, followed by treatment with 5 μM of Aβ(1–42) with or without TA (10 μM) for 24 h. After treatment, the BV-2 cells were fixed and examined by a Leica SP8 confocal microscope with a LAS X 3D Visualization (Leica Microsystems Inc., Wetzlar, Germany). Representative images of BV-2 cells with co-localization of pmCherry-C1-ASC and GFP-LC3 puncta were captured, and the image graining was performed by the Leica SP8 processor. Magnification, × 63; scale bar: 25 μm. Error bars, S.D. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001, n=6. (One-way ANOVA with Tukey-corrected post hoc t-tests for multiple comparisons was applied for comparison between groups). The full-length blots are presented in Additional file 1: Fig. S21, 22