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. 2022 Oct 21;13:6275. doi: 10.1038/s41467-022-34005-1

Fig. 6. PU-H71 treatment reduces oTau-induced pathology and neurodegeneration.

Fig. 6

a Experimental schematic including 2D culture, oligomer exposure, 3D culture, PU-H71 treatment, and analysis at indicated timepoints. Created with BioRender.com. b Representative images showing reduction in tau misfolding in AstTau upon exposure to PU-H71 for 3 days, from 19 to 21 DIV3D. Misfolded tau (MC1, green), neurons (βIII tubulin (Tuj1), violet), astrocyte (S100β, red). Scale bars = 20 μm. c Quantification of MC1 labeled fluorescence intensity normalized to Tuj1 intensity. Data obtained from 5 independent asteroids. Error bars = SEM. Two-way ANOVA with Tukey’s multiple comparisons test was performed, ****P < 0.0001 comparisons to timepoint vehicle control. d Representative images showing reduction in ThioS (fibrillar tau), in AstTau upon exposure to PU-H71 for 3 days, from 19 to 21 DIV3D. Misfolded tau (MC1, green), neurons (Tuj1, violet), astrocyte (S100β, red) and (GFAP, blue). Scale bars =50 μm. e, f Quantification of ThioS (e) and S100B (f) labeled fluorescence intensity normalized to Tuj1 (e) or DAPI (f) intensity. Data obtained from five independent asteroids. Error bars = SEM. Two-way ANOVA with Tukey’s multiple comparisons test was performed, ****P < 0.0001 comparisons to vehicle control at each timepoint. g Representative images showing Fluoro Jade B-labeled fluorescence in AstTau upon exposure to PU-H71 for 3 days, from 19 to 21 DIV3D. Damaged neurons are labeled by Fluoro Jade B (green). Neurons (Tuj1, violet), astrocyte (S100β, red) and (GFAP, blue). Scale bars = 50 μm. h Quantification of Fluoro Jade B-labeled fluorescence intensity normalized to Tuj1 intensity. Data obtained from five independent asteroids. Error bars = SEM. Two-way ANOVA with Tukey’s multiple comparisons test was performed, ****P < 0.0001 comparisons to vehicle control at each timepoint. i LDH detection in the conditioned medium of 21 DIV3D asteroids. Data obtained from 25 independent asteroids. Error bars = SEM. One-way ANOVA with Tukey’s multiple comparisons test was performed, **P < 0.01.