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. 2019 Apr 24;13:152. doi: 10.3389/fncel.2019.00152

Figure 8.

Figure 8

P2Y1 is necessary to maintain myosin light chain (MLC) phosphorylation at the AIS. (A) Representative images of 7 DIV scrambled shRNA or P2Y1 shRNA nucleofected neurons showing phosphorylation of MLC (pMLC) staining in the AIS. Neurons were stained with MAP2 (blue) and pMLC (red) antibodies. Nucleofected neurons were identified by GFP signal (green). pMLC staining at axon initials segments was magnified and presented in panels below (red). (B) Scatter plot for each neuron (left) and mean ± SEM (right) representing normalized pMLC intensity at the AIS of 7 DIV neurons nucleofected with scrambled shRNA or P2Y1 shRNA. ***p < 0.001, two-tail t-test. (C) Scatter plot representing normalized pMLC intensity at the AIS of 7 DIV neurons nucleofected with scrambled shRNA or P2Y1 shRNA and treated with 1 nM calyculinA (CalyA) for 3 h before fixation. Mean ± SEM of these values is represented in the adjacent graph. (D) Scatter plot representing normalized ankyrinG intensity at the AIS of 7 DIV neurons nucleofected with scrambled shRNA or P2Y1 shRNA and treated with 1 nM calyculinA (CalyA) for 3 h before fixation. Mean ± SEM are represented in the adjacent graph. Statistical differences in (C,D) were analyzed by a Kruskal-Wallis test followed by a Dunn’s multiple comparisons post-test. *p < 0.05, **p < 0.01, ***p < 0.001.