Fig. 4

LRP4 LDLα domain increases the dendrites arborization in primary culture neurons. a Schematic diagram of time course dealing with the primary culture neurons. pFLAG-CMV1-Lrp4-LDLα, pFLAG-CMV1-Reelin, and pFLAG-CMV1 vector plasmids were transfected into HEK293T cells, and then the conditioned medium was collected to add into primary cultured wild-type mice neurons, respectively. In panel B-D, pFLAG-CMV1 vector group, pFLAG-CMV1-Lrp4-LDLα group, and pFLAG-CMV1-Reelin group were abbreviated as control, LRP4 LDLα and REELIN, respectively; b Representative images of neurons staining in primary culture; Using the ImageJ plugin tracing the neurite branches; c Western blotting check the expression of the plasmids after transfected into HEK293T cells. Anti-Flag for LRP4 and REELIN immunoprecipitation from the cell culture medium. The input was from the cell lysis; d LRP4 LDLα domain improved the number of dendritic branches in primary neurons. The ImageJ plugin was used for Sholl analysis. Multiple comparisons with two-way ANOVA analyses were made between the LRP4 LDLα group and the control (between the curves). (Scale bar = 30 μm; Wild-type mice, n = 8; Neuron number, control n = 11, LRP4 LDLα n = 10 and REELIN n = 13. Values are means ± SEM. * P < 0.05, ***P < 0.001, *** P < 0.001)