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. 2018 Jun 8;64(1):205–221. doi: 10.3233/JAD-180054

Fig.3.

Fig.3

Pyk2 phosphorylates tau in HEK293T cells. A) Representative immunoblots of lysates from HEK293T cells transfected with V5-tagged tau and GFP-tagged Pyk2. Validation of the Y402F mutant form of Pyk2 by immunoblotting using a phospho-Y402-Pyk2 specific antibody. Sh, short exposure; lo, long exposure. Asterisk denotes the endogenous expression of Pyk2 in HEK cells using a longer exposure of the immunoblot in the absence of Pyk2-GFP overexpression. B) Quantification of immunoblots in (A) showing relative tau and pY18-tau levels. Mean±s.e.m, n = 6 per group, one-way ANOVA, Dunnett’s multiple comparisons test, **p < 0.01, ****p < 0.0001. C) Representative immunofluorescence staining of HEK293T cells transfected with V5-tagged tau and GFP-tagged Pyk2 indicating a strong positive correlation of pY18-tau immunoreactivity and expression of the active form of Pyk2. Scale bar: 50μm. D) Immunoblots of lysates from HEK293T cells co-transfected with V5-tagged tau and PTK2B or control siRNA. E) Quantification of immunoblots in (D). Mean±s.e.m, n = 3 per group, two-tailed t test, *p < 0.05, ***p < 0.001.