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Fig. 2 | Neural Development

Fig. 2

From: Precise levels of nectin-3 are required for proper synapse formation in postnatal visual cortex

Fig. 2

Nectin-3 knockdown at E15.5 increases dendritic spine density at P21. a Co-electroporated expression constructs. b Top: Western blot showing nectin-3 expression after co-transfection of HEK-293 cells with a nectin-3 expression plasmid and either nectin-3 or scramble shRNA constructs. Bottom: Nectin-3 expression/intensity was quantified and normalized to α-tubulin. Expression is shown relative to average nectin-3 expression from the scramble shRNA condition. c Electroporation/shRNA expression occurred at E15.5. Dendritic spine densities were analyzed at P14, P21, and P35. d Electroporated neurons at P21 in visual cortex (10x image, scale bar = 500 μm). Inset top: Cells expressing Cre-dependent tdTomato did not express floxed GFP-stop from pSico (40x image, scale bar = 50 μm). Inset bottom: Representative V1 neuron (40x image, scale bar = 50 μm), and select apical and basal dendrites. e Differences in dendritic spine densities between control and Nec3-shRNA neurons. Error bars denote standard errors of the mean. From left to right, asterisks denote significant differences for pooled apical and basal dendrites (solid line) between time points or between conditions at P21 (Additional file 5). f Representative apical dendrites from P21 Nec3-shRNA and control neurons (63x image, scale bar = 10 μm). g Nec3-shRNA and control dendritic spine densities at P14. Apical and basal dendritic spine densities are plotted. Error bars denote standard error of the mean. Each dot represents a single dendrite. One apical and one basal dendrite per cell were counted (Control: N = 15 cells, 30 dendrites, 5 animals; Nec3-shRNA: N = 16 cells, 32 dendrites, 6 animals). h Nec3-shRNA and control dendritic spine densities at P21. Significance is denoted by ‘*’ (Control: N = 17 cells, 34 dendrites, 5 animals; Nec3-shRNA: N = 14 cells, 28 dendrites, 5 animals). i Nec3-shRNA and control dendritic spine densities at P35 (Control: N = 11 cells, 22 dendrites, 3 animals; Nec3-shRNA: N = 10 cells, 20 dendrites, 5 animals)

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