Fig. 4: March5 promotes Kindlin-2 protein levels by inhibiting Trim28.

a IF staining. INS1 cells were subjected to IF staining with an anti-March5 antibody (green) and an anti-Trim28 antibody (red). Scale bar, 10 μm. N = 3 biologically independent samples. b, c Co-IP assays. INS1 cells were transiently transfected with plasmids expressing Kindlin-2 or Trim28, and the cell lysates were collected for IP and IB with the indicated antibodies. N = 3 biologically independent samples. d–g IHC staining for Trim28. Pancreatic sections from CON and cKO, CON and M5OE mice were subjected to IHC staining for Trim28. Scale bar, 50 μm. Quantitative data from 5 mice per group (f, g). h–k Western blot analyses. HEK293T cells were transiently transfected with the indicated plasmids for 48 h. Cell lysates were then subjected to western blot analyses with the indicated antibodies. Quantitative data from three independent experiments (i, k). l IF staining. INS1 cells were subjected to IF staining with an anti-March5 antibody (red), an anti-Trim28 antibody (green), and DAPI (blue). Scale bar, 10 μm. N = 3 biologically independent samples. m Co-IP assays. INS1 cells were transiently transfected with plasmid expressing Trim28 for 48 h, the cell lysates were then collected for IP and IB with the indicated antibodies. N = 3 biologically independent samples. Data are expressed as mean ± s.d. Data were analyzed by two-tailed unpaired Student’s t-tests for comparisons between two groups and one-way ANOVA for multiple groups, followed by Tukey’s post hoc test. Source data are provided as a Source Data file.