Analysis of selected STAB2 missense mutants. Reference STAB2 cDNA or cDNA encoding 7 different missense variants was used to establish stable stabilin-2 expressing cell lines in flp in TREx 293 cells. (A) On-Cell Western quantification of stabilin-2 surface expression was performed using an anti-ectodomain antibody and quantified with a Li-Cor Odyssey CLx and Image Studio software. IR antibody signal was normalized to cell counts, and reference sequence was mean centered to 100. All VTE variants had lower mean surface expression (P < .0001, except G1597S, P = .0014, 2-tailed Student t test). (B) Flow cytometry on live cells using an antibody against the receptor ectodomain to estimate median fluorescent signal corresponding to the amount of stabilin-2 receptor on the surface of the cells. (C) Cell lysate was obtained and incubated with and without Endo H and then western blotted using fluorescently conjugated secondary antibodies. Bands were quantified using image studio software, and the ratio of undigested to total bands was plotted for each mutant. For panels B and C, P values are reported in comparison of missense variant to ref STAB2. *P < .05, **P < .005, ***P < .0005, ****P < .0001 by 2-tailed Student t test.