Activation of Rac1 and RhoA, especially Rac1, played a central role in podocyte actin filament rearrangement induced by Angptl3. (a), (b), and (c) Changes in the activation of small GTPases Rac1, RhoA, and Cdc42 after treatment with Angptl3 (250 ng/mL) for different times. The relative total small GTPase protein levels were detected by western blot and are shown in the top panel. The levels of the active forms of the small GTPases were detected with G-LISA and are shown in the bottom panel. No lysate was added in negative control groups, while control protein instead of lysate was added in positive control groups. Compared to the control group, *, **. Compared to the peak value, #, ##. (d) Lamellipodia formation in podocytes after blocking Rac1, followed by Angptl3 treatment. (A) F-actin in podocytes incubated with 80 mM NSC23766 for 2 h. (B) F-actin in podocytes preincubated with 80 mM NSC23766 for 2 h, followed by 250 ng/mL Angptl3 treatment for 30 min. (C) F-actin in podocytes preincubated with 80 mM NSC23766 and 80 mM CT04 for 2 h, followed by 250 ng/mL Angptl3 treatment for 30 min. Scale bar, 20 μm. The comparison of lamellipodia formation between groups (A) and (B) is shown in (e).
