Journal: Scientific Reports

Article Title: Mitochondrial sub-cellular localization of cAMP-specific phosphodiesterase 8A in ovarian follicular cells

doi: 10.1038/s41598-019-48886-8

Figure Lengend Snippet: Immunolocalization in mitochondrial isolated fractions from granulosa cells by immunofluorescence using anti-P450scc (green, B , United States Biological), anti-PDE8A (green, E , Proteintech) and MitoTracker (orange, A , D ). Merge signals are shown in ( C , F ). The signal was calibrated with non-specific IgG. Magnification is shown as a bar of 10 µm.

Article Snippet: The samples were washed twice in PBS solution, centrifuged at 14,000 × g for 5 min to eliminate residual markers, fixed with 4% (w/v) PFA in 0.2% Triton X-100 for 10 min in the dark at room temperature, blocked with 5% BSA in PBS for 1 h, then kept at 4 °C overnight with PDE8A polyclonal antibody (Proteintech product number 13956-1-AP, 1:500 in PBS) or porcine P450scc polyclonal antibody (United States Biological, Salem, MA, product number 362774, 1:100 in PBS).

Techniques: Isolation, Immunofluorescence