Wnt5a acts through a non-canonical pathway and decreases the number of presynaptic inputs. (A, B) Representative images of 'active' β-catenin immunostaining in control (A) and Wnt5a (A; 200 ng/mL) treated cells. Scale bar = 6 μm. (C, D) Quantification of 'active' β-catenin immunofluorescence intensity (± standard error of the mean) in the nucleus (C) and soma (D). Wnt5a treated cultures were normalized to the control condition (n = 220 cells, ***p < 0.001, Students t-test). (E) Representative images of control (left) and Wnt5a (right; 200 ng/mL) treated cells immunostained with Vesicular glutamate transporter 1 (VGLUT1; green) and Microtubule associated protein 2 (MAP2; dark blue) at 10 days in vitro (10 DIV). Scale bar = 2 μm. (F) Quantification of VGLUT1 puncta per area of dendrite (± standard error of the mean; n = 60 fields of view; **p = 0.001, Students t-test). (G) Cultures in which endogenous Wnts were inhibited with secreted Frizzled related protein 2 (sFRP2). Control (left) and sFRP2 (right; 200 ng/mL) treated cells were stained at 14 DIV with VGLUT1 (green) and MAP2 (dark blue). Scale bar = 2 μm. (H) Quantification of sFRP2 effects on the number of VGLUT1 puncta per area of dendrite (± standard error of the mean; n = 59 fields of view; *p = 0.05, Students t-test).