Fig 1.

Activation of Erk1/2 and transcription factors CREB and ATF-1 by DHEAS in TM4 cells detected by immunofluorescence: TM4 cells were incubated with or without 1 μM DHEAS for 120 min and then stained with specific antibodies against either phospho-Erk1/2 (A-B), phospho-CREB (D-E) or phospho-ATF-1 (G-H) and a secondary Alexa Fluor 488-labeled antibody as described under “Methods”. Green fluorescence indicates the phosphorylation of Erk1/2 (A-B), CREB (D-E) of ATF-1 (G-H). Nuclei of the cells were labeled with DAPI and appear blue. (A, D and G) Fluorescence of phospho-Erk1/2, phospho-CREB or phospho-ATF-1 in the absence of DHEAS. (B, E and H) Phosphorylation of Erk1/2, CREB and ATF-1 in response to DHEAS (1 μM) treatment. (C, F and I) Statistical analysis of the corresponding green fluorescence in arbitrary units (n = 90; means ±SEM; **p≤0.01).