Figure 1

FOXO3 induces DEPP expression on mRNA and protein level. a) Regulation of DEPP (probe set 209183_s_at) as measured by HG-U133 Plus 2.0 microarrays. Shown are fold-increases in expression between 3 hours 4OHT treated and untreated samples. b) Quantitative RT-PCR of DEPP expression in SH-EP/FOXO3, NB1/FOXO3, NB3/FOXO3 and NB15/FOXO3 cells treated for 0, 3 and 6 hours with 100 nM 4OHT. Shown are means ± s.e.m. of three independent experiments, each performed in triplicates. c) Immunoblot analyses of DEPP expression in SH-EP/FOXO3, NB1/FOXO3, NB3/FOXO3 and NB15/FOXO3 cells treated with 100 nM 4OHT for 0, 4, 8, 16 and 24 hours. Alpha-Tubulin served as loading control. d) Quantitative RT-PCR of DEPP expression in SH-EP/Ctr and SH-EP/FOXO3-DBD cells, which were cultured in 0.5% FCS for 24 hours and then transferred into standard media (10% FCS) or treated with 10 nM insulin for another six hours. Shown are mean values ± s.e.m of three independent experiments, each performed in triplicates; statistical analysis was done with the Student’s unpaired t-test, *P < 0.05; ***P < 0.01 compared to corresponding controls. e) Immunoblot analysis of DEPP expression in SH-EP/Ctr cells, which were cultured in 0.5% FCS for 24 hours. GAPDH served as loading control.