Extended Data Fig. 9: Manipulation of Rho GTPase regulators alters synapse development in human and mouse cortical neurons.

a, Original large field of view images for producing processed images in Fig. 4c (scale bar: 50 μm). b, Immunostaining of dendrites from primary mouse cortical neurons cultured 8 days in vitro. Neurons were transfected with mEGFP-C1 and vectors expressing mCherry, mCherry-ARHGEF7, or mCherry-RASGRF2 (n = 20, 20 and 20 neurons from 4 cultures, scale bar: 5 μm). Data are presented as mean values ± s.e.m. The P values were obtained from one-way ANOVA with Holm-Sidak’s multiple comparisons test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. c, Original large field of view images for producing processed images in panel b (scale bar: 50 μm). d, Quantification of mRNA levels of ARHGAP23 and SRGAP1 in HEK293T cells transfected with control shRNAs (shControl), two shRNAs targeting ARHGAP23 (shARGGAP23−1 and shARGGAP23−2), or two shRNAs targeting SRGAP1 (shSRGAP1−1 and shSRGAP1−2) (n = 3 cultures for all conditions). Data are presented as mean values ± s.e.m. The P values were obtained from one-way ANOVA with Holm-Sidak’s multiple comparisons test; ****P < 0.0001. e,f, Immunostaining of dendrites from primary human cortical neurons cultured six weeks in vitro. Neurons were transfected with mEGFP-C1 and vectors co-expressing turbo-RFP (tRGP) and shControl, shARGGAP23−1, shARGGAP23−2, shSRGAP1−1 or shSRGAP1−2 (n = 20, 22, 20, 20 and 20 neurons from 4 cultures, scale bar: 5 μm). Data are presented as mean values ± s.e.m. The P values were obtained from one-way ANOVA with Holm-Sidak’s multiple comparisons test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. g, Original large field of view images for producing processed images in panel e (scale bar: 50 μm). h, Original large field of view images for producing processed images in Fig. 4e (scale bar: 50 μm).