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. 2017 Oct 3;21(1):84–96. doi: 10.1016/j.celrep.2017.09.019

Figure 4.

Figure 4

GRIP1 Is Required for Activity-Induced Dab1 Phosphorylation Mediated by ApoER2

(A and B) Dab1 phosphorylation in response to neuronal activity was examined in primary hippocampal neurons prepared from wild-type (+/+) and GRIP1 knockout (GRIP1−/−) embryos at E15.5. Images of immunocytochemical pDab1 staining in dendrites stimulated with 10mM KCl for 10 min (A). Quantification of pDab1 fluorescence intensities is shown (n = 3) (B).

(C) Single-heterozygous neurons for ApoER2 and GRIP1 show normal levels of pDab1. Quantification of relative pDab1 fluorescence intensities in wild-type, ApoER2+/−, and GRIP1+/− neurons (n = 3–6).

(D and E) ApoER2 and GRIP1 genetically interact in the phosphorylation of Dab1 upon KCl stimulation. Primary hippocampal neurons from wild-type and ApoER2+/−; GRIP1+/− compound mice were stimulated with KCl, and pDab1 levels were assessed by immunocytochemistry. Fluorescent images represent pDab1 in wild-type (+/+) and ApoER2+/−; GRIP1+/− compound neurons (D). Quantification of relative pDab1 fluorescence intensity in wild-type and ApoER2+/−; GRIP1+/− compound neurons is shown (n = 3) (E).

Scale bars represent 20 μm in (A) and (D) and 5 μm in the higher magnifications in (A) and (D). Bar graphs show mean ± SEM (shown as error bars). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.