GSK3α and GSK3β knockdown alter LatrB-induced changes to dendritic spine morphology.

<p>A. GSK3 α/β knockdown does not affect basal fluctuations of dendritic spine morphology. Experimental outline with 4 time points for microscopy and quantitative analysis of spine shape; # indicates <i>p</i><0.05 for measurements of spines after GSK3β silencing compared to control values at the corresponding time points. For number of counted spines refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134018#pone.0134018.t003" target="_blank">Table 3</a>. Data are presented as the mean spine width per cell ± s.e.m. The curve between time points is extrapolated. B. Experimental outline with 3 time points for microscopy: baseline, LatrB treatment, end of recovery period. Representative micrographs of cultured DIV18 murine hippocampal neurons transfected with shRNA constructs as indicated. Scale bar = 2.5 μm. C. Quantitative analysis of spine shape changes; *** = <i>p</i> < 0.001 and xx = <i>p</i> < 0.01, xxx = <i>p</i> < 0.001 and ### = <i>p</i> < 0.001 for spine measurements of shRNA silenced GSK3α, GSK3β and GSK3α/β compared to the control at the corresponding time points. ‡ = <i>p</i> < 0.05 difference between shRNAGSK3α and shGSK3β. For number of counted spines refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134018#pone.0134018.t003" target="_blank">Table 3</a>. Data are presented as mean spine width per cell ± s.e.m. The curve between time points is extrapolated. D. Spine l/w ratio changes presented as cumulative histograms of length/width ratio at 3 time points.