Fig. 5
CARMIL3 depletion causes alterations in dendritic spine maturation. a Schematic of CRISPR-based depletion strategy of CARMIL3 in cultured hippocampal neurons. b Representative images of control (WT) and CARMIL3-depleted (MUT) neurons at DIV8. Scale bars are 5 μm. c Graphical representation of dendritic spine density for WT (18 ± 1 spines, n=28 neurons) and MUT (18 ± 2 spines, n=28 neurons) neurons at DIV8. p=0.8235. d Graphical representation of filopodia density for WT (14 ± 1 filopodia, n=28 neurons) and MUT (13 ± 1 protrusions, n=28 neurons) neurons at DIV8. p=0.4541. e Representative images of WT and MUT neurons at DIV16. Scale bars are 5 μm. f Graphical representation of dendritic spine density for WT (48 ± 2 spines, n=29 neurons) and MUT (24 ± 2 spines, n=29 neurons) neurons at DIV16. p<0.0001. g Graphical representation of dendritic filopodia density for WT (13 ± 1 filopodia, n=29 neurons) and MUT (25 ± 2 filopodia, n=29 neurons) neurons at DIV16. p<0.0001. h Schematic of HITI gene trap strategy utilized for depleting CARMIL3 in vivo. i Representative images of tdTomato fill in WT and MUT pyramidal neurons at postnatal day 14 (P14) in hippocampal CA1. Scale bars are 5 μm. j Graphical representation of dendritic protrusion density for WT (81 ± 5 protrusions, n=20 neurons) and MUT (62 ± 5 protrusions, n=11 neurons) neurons at P14. p=0.018. k Graphical representation of dendritic spine density for WT (74 ± 5 spines, n=20 neurons) and MUT (53 ± 6 spines, n=11 neurons) neurons at P14. p=0.018. l Graphical representation of dendritic filopodia density for WT (7 ± 1 filopodia, n=20 neurons) and MUT (9 ± 2 filopodia, n=11 neurons) neurons at P14. Error bars are standard error of the mean (SEM). p=0.272. *p<0.05, **p<0.01, ****p<0.0001, t-tests
