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. 2021 Jan 4;26(6):1775–1789. doi: 10.1038/s41380-020-00991-1

Fig. 3. Mutation of the PPXXF binding motif in ankyrin-G 190 attenuates its interaction with Homer1b/c in dendrites.

Fig. 3

a SIM images to detect the interaction between HA-ankyrin-G WT or mutants (P1606L or F1609R) and anti-Homer1b/c by in situ PLA. The PLA was conducted after transfection of HA-ankyrin-G WT or mutant constructs into the cultured mouse cortical neurons. Negative control was transfected with only mCherry construct. Scale bar, 2 µm. b The number of PLA puncta per 10 µm from HA-ankryn-G WT, P1606L, and F1609R transfected neurons. PLA was performed with anti-Homer1b/c and anti-HA antibodies. F(2, 123) = 11.75. c Spine head size from mushroom spines was analyzed from only mCherry or HA-ankryn-G WT, P1606L, and F1609R transfected neurons with mCherry. F(3, 1445) = 18.62. d Total numbers of all spines were counted. F(3, 141) = 3.64. e Mushroom, thin, and stubby types of spine morphology were analyzed. F(6, 423) = 5.59. n = 13 (only mCherry), 17 (WT), 19 (P1606L), 21 (F1609R) cells. *p < 0.05; **p < 0.01; ***p < 0.001; one-way ANOVA followed by a Bonferroni test for b–d and two-way ANOVA followed by a Bonferroni test for e. Data are represented as mean ± SEM.