Figure 3. Lztr1 loss affects vesicular trafficking.

(a) IPA analysis of differentially expressed proteins in HeLa cells harboring wt-LZTR1 or LZTR1-indels. Differentially expressed proteins were identified by MS-based shot gun analysis using the FDR approach. N=3 (b) IPA analysis of differentially ubiquitinated peptides in HeLa cells harboring wt-LZTR1 or LZTR1-indels. GG-modified peptides were captured by anti-K-ε-GG antibody and quantified by MS. Differentially ubiquitinated proteins were identified by the FDR approach. N=3 (c) Putative LZTR1 interactors identified by the Virotrap approach as described in Steklov et al., 2018. Endocytosis-related proteins are shown in red. N=3 (d, e) Lztr1 ^fl/fl ECs expressing an empty vector (EV) or Cre recombinase were immunostained with anti-SNX1 antibody. Scale bar 10 μm. Each dot shows mean value for 25 ECs isolated from one mouse. Lines show mean±SEM; p-values were assessed by Wilcoxon-Mann-Whitney test. Sphericity ranges from 0 to 1 indicating if the structures are spheric (close to 1) or more tubular (close to 0.5). (f, g) Lztr1 ^fl/fl ECs transduced with an empty vector (EV) or Cre-recombinase coding virus (Cre) were treated with DMSO, Dynasore (40μM), or Tyrphostin A23 (10μM) for 12 hours and immunostained with anti-ZO1 and anti-VE-cadherin antibody. Scale bar 20 μm. Quantification of the number of junctional gaps per cells. N=4; 25 cells per group were analyzed; p-values were assessed by Kruskall Wallis test with Dunn’s Multiple Comparison Test (h) Progeny from Lztr1^+/− matings at E18.5 and after birth in a mock group or a dynasore-treated group (1mg/kg; starting from E2.5). P-values were calculated using Fisher Exact test.