Fig 1: ASAP1 is increased in endothelial cells of diabetic mice and negatively regulates ARF4 activity. a) ASAP1 expression in mice kidney cells by single‐cell sequencing analysis of kidneys from diabetic OVE26 mice and control mice. b) Representative ASAP1 immunofluorescence images and quantification of mean glomerular ASAP1 intensity per mouse (n = 5 mice per group, 20–30 glomeruli scored per mouse). Scale bar = 20 µm. c) HUVECs were treated with normal glucose (NG) or high glucose (HG) and were immunoprecipitated with GGA3 beads and immunoblotted for ARF4. The top panel shows the immunoprecipitated proteins and the bottom panel shows total input. d) HUVECs were infected with Flag‐tagged ASAP1 or control vectors and then treated with either vehicle or 1 nm 25‐HC. Cells were immunoprecipitated with GGA3 beads and immunoblotted for ARF4. The top panel shows the immunoprecipitated proteins and the bottom panel shows total input. e) Measurement of Gaussia luciferase activity in the medium of HUVECs overexpressed with or without ASAP1 and treated with or without 25‐HC. f) Representative images and quantification of proximity ligation assay (PLA) between ASAP1 and ARF4 in HUVECs treated with or without 25‐HC. *P < 0.05, **P < 0.01 by 2‐way ANOVA. ****P < 0.0001 by t‐test. ns, non‐significant.