Skip to main content
. 2022 Jan 1;12(3):1303–1320. doi: 10.7150/thno.67702

Figure 4.

Figure 4

Rac1 mediates Hh signaling by control of stabilization of IFT88. (A) Immunoblotting analyses of IFT88 in C3H10T1/2 cells transfected with or without daRac1 and cultured for 24 h. (B) Immunoblotting analyses of IFT88 in C3H10T1/2 cells cultured with or without NSC23766 at 10 μg/ml for 24 h. (C) Immunoblotting analyses of IFT88 in C3H10T1/2 cells transfected with or without daRac1 and treated for different time periods with cycloheximide (CHX). (D) Immunoblotting analyses of IFT88 in C3H10T1/2 cells cultured with or without NSC23766 (NSC) at 10 μg/ml for 24 h and treated for different time periods with CHX. (E) Immunoblotting analyses of IFT88 in C3H10T1/2 cells cultured with or without NSC23766 at 10 μg/ml and MG132 at 10 μM for 24 h. (F) Immunofluorescence staining for IFT88 in MEFs with or without NSC23766 at 10 μg/ml for 24 h. Primary cilia were indicated by Arl13b staining. Nuclei were counterstained by DAPI. Bar, 20 μm. (G) C3H10T1/2cells were transiently transfected with a Gli luciferase reporter together with shIFT88 for 48 h and daRac1 for 24 h. Total cell lysates were subjected to luciferase assay. N=6. (H) C3H10T1/2 cells were transfected with shIFT88 and daRac1. Total cell lysates (Input) and anti-SuFu immunoprecipitates (IP) from total cell lysates were analyzed by immunoblotting with anti-Gli1 and anti-SuFu antibodies. IgG was used as a negative control for IP. (I) Statistical analysis for the percentage of ciliated C3H10T1/2 cells transfected with or without shIFT88. N=6. (J) Statistical analysis for the percentage of ciliated C3H10T1/2 cells cultured with or without NSC23766 (NSC) at 10 μg/ml for 24 h. N=6. Protein abundance normalized to GAPDH or αTubulin, respectively. **, ##p < 0.01; error bar, SD.