Journal: The Journal of Biological Chemistry
Article Title: Plasma Membrane Origin of the Steroidogenic Pool of Cholesterol Used in Hormone-induced Acute Steroid Formation in Leydig Cells *
doi: 10.1074/jbc.M116.740928
Figure Lengend Snippet: Cholesterol trafficking analysis in primary rat Leydig cells infected with lentivirus containing mCherry-D4. A, confocal microscope images of cells with and without addition of Bt2cAMP. Scale bar, 10 μm. B, time course of mCherry-D4 fluorescence intensity at the plasma membrane of control and Bt2cAMP-treated cells. C, testosterone content of control cells and cells infected with lentivirus before and after a 2-h treatment with Bt2cAMP. Testosterone was measured just after the cells were isolated (0 h), in control (uninfected) cells 18 h after incubation and 18 h after infection. Data represent means ± S.D. of at least three independent experiments performed in triplicate; two-way ANOVA followed by Bonferroni's post hoc test (*) was used to calculate statistical significance. *, p < 0.05; **, p < 0.01; ***, p < 0.001. dbcAMP, dibutyryl cAMP.
Article Snippet: MA-10 cells from mouse Leydig cell tumor (kindly provided by Dr. M. Ascoli, University of Iowa, Iowa City) and R2C cells from rat Leydig cell tumor cells (American Type Culture Collection, Manassas, VA) were cultured with Dulbecco's modified Eagle's medium (DMEM)/Ham's F-12 (Invitrogen) supplemented with 1% penicillin and 1% streptomycin and maintained at 37 °C.
Techniques: Infection, Microscopy, Fluorescence, Isolation, Incubation