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. Author manuscript; available in PMC: 2022 Aug 19.
Published in final edited form as: Cell. 2021 Jul 26;184(17):4547–4563.e17. doi: 10.1016/j.cell.2021.07.003

Figure 6. ELAVL4 binds MAPT RNA and co-localizes with cytosolic stress granules in tau-V337M neurons.

Figure 6.

(A) ELAVL4 RNA immunoprecipitation (RIP) detects MAPT 3′ untranslated region (UTR) and exon 13 in tau-V337V (left panel) and tau-V337M organoids (right panel). CALM3, positive control.

(B) Expression of TIA1 and G3BP1 in tau-V337M and tau-V337V glutamatergic neurons. Dot size, proportion of cells expressing a gene; depth of color, expression level. Values: differential gene expression p value adjusted by MAST general linear model.

(C and D) G3BP1 immunostaining (C) and quantification of G3BP1 intensity relative to DAPI (D) in tau-V337V and V337M organoids at 2 months. Bars represent mean intensity ± SD. Unpaired t test, *p ≤ 0.01. n = 4 images per organoid; n = 3 organoids per line for 2 independent experiments.

(E and F) Western blot and densitometry quantification of G3BP1 in tau-V337M and isogenic V337V organoids at 2 months. Bars represent G3BP1 densitometry in mutant organoids (%) relative to isogenic controls ± SEM. Unpaired t test, **p ≤ 0.01; n = 6 per group for 2 independent experiments.

(G) ELAVL4 and G3BP1 colocalization (white arrows) in tau-V337V and V337M organoids at 2 months. Scale bar, 20 μm.

(H) ELAVL4 and TIA1 co-localization with tau at 2 months (scale bar, 5 μm) and ELAVL4 with tau at 4 months (white arrows; scale bar, 10 μm).

(I) Co-localization of tau, ELAVL4, and G3BP1 in tau-V337M organoids at 4 months. Scale bar, 5 μm.