figs2.

Secretagogin increases Ngn3 protein level without affecting Ngn3 promoter activity. (A) Ectopic expression of Scgn (CMV-Scgn; 48 h) in HEK293T cells does not affect the activity of the Ngn3 promoter. Promoter activity was defined as the ratio of firefly-to-Renilla luciferase chemiluminescence. (B–B₁) HEK293T cells where transfected with Ngn3-expression constructs (CMV-Ngn3). Scgn co-expression (CMV-Scgn; 48 h) significantly increased Ngn3 protein levels as shown by Western blotting. Rescuing Ngn3 degradation by proteasome inactivation through lactacystin (5 μM, 6 h) was used as positive control. Representative immunoblot is shown. Total protein load (Cy5) was used to normalize Ngn3 protein levels. Data were expressed as means ± s.d. from triplicate experiments, *p < 0.05 (Student's t-test in A or one-way ANOVA in B–B₁).2