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. 2021 Nov 29;27:256–275. doi: 10.1016/j.omtn.2021.11.019

Figure 6.

Figure 6

PTEN/p38 MAPK signaling contributes to the inhibitory effects of miR-148a-3p on tau phosphorylation

(A and B) Representative western blot images of phosphorylation of tau protein at AT8, S199, S396, and S404 sites and Tau (A) and quantification of the ratios of AT8/Tau, S199/Tau, S396/Tau, and S404/Tau (B) after transfection of APPswe cells with PTEN-expressing plasmid (PTEN), vector control (Vector), PTEN siRNA, and scrambled siRNA (Scr siRNA). (C) Co-localization of PTEN with PHF-1, the Ser396/404 sites of tau phosphorylation, and total tau (Tau) in APPswe cells using an immunofluorescence assay. Scale bar, 50 μm. (D) Quantitative analysis of co-localization of PTEN with PHF-1 and Tau using Pearson coefficient in Fiji ImageJ software. (E and F) Representative western blot images of p-PTEN, PTEN, pTS396, pTS404, Tau, p-p38 MAPK, p38 MAPK, p-ERK1/2, ERK1/2, p-GSK3β, and GSK3β (E) and quantification of PTEN expression and ratios of p-PTEN/PTEN, S396/Tau, S404/Tau, p-GSK3β/GSK3β, p-ERK1/2/ERK1/2, p-JNK/JNK, and p-p38 MAPK/p38 MAPK (F) after transfection of APPswe cells with PTEN-expressing plasmid (PTEN), vector control (Vector), PTEN siRNA, and scrambled siRNA (Scr siRNA). (G and H) Representative western blot images of phosphorylation of tau protein at AT8, S199, S396, and S404 sites; total tau; p-PTEN; and PTEN (G) and quantification (H) after transfection of APPswe cells with PTEN-overexpressing plasmid (PTEN) and PTEN-expressing plasmid plus miR-148a-3p mimic (PTEN+148a). Results represent means ± SEM, n = 5. ∗n < 0.05, ∗∗n < 0.01 versus vector or Scr siRNA or PTEN + Tau. $p < 0.05, $$p < 0.01, $$$p < 0.001 versus PTEN. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.