Figure 4: Net4 blocks laminin assembly on the cell surface.
From: Structural decoding of netrin-4 reveals a regulatory function towards mature basement membranes
(a) Within untreated (ctrl), Net4-FL-, and Net4-FLE195A,R199A-treated (28-fold excess in molarity compared with laminin) Schwann cell extracellular matrix assembly assay BM components collagen IV (Col-IV, green) and laminin 111 (Lmγ1, red) were stained. Cells were counter-stained for DAPI (blue). (b) Staining intensities are displayed relative to the DAPI signal [relative Lmγ1 intensity per cell (left) and relative Col-IV intensity per cell, (right)] (mean±s.d.; n=5; ****P<0.00001; ns, not significant). Error bars, s.d. (n=5 independent cell cultures). (c) Concentration dependent effect of Net4-FL and Net4-FLE195A,R199A on BM formation indicated through measurement of the relative Lmγ1 intensity/cell (left) and relative Col-IV intensity/cell (right). Net4 proteins were added together with the BM components laminin 111, collagen IV and nidogen-1 with increasing molar ratios to laminin 111 (0.4, 1.8, 7, 28-fold excess). Scale bars, (a) 200 μm. P values were calculated by one-way ANOVA followed by a pairwise comparison using the Holm-Sidak method. ANOVA, analysis of variance.
