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Fig. 2 | Reproductive Biology and Endocrinology

Fig. 2

From: Docosahexaenoic acid (DHA) effects on proliferation and steroidogenesis of bovine granulosa cells

Fig. 2

Expression and localization of free fatty acid receptor 4 (FFAR4) in bovine granulosa cells by immunofluorescence. Immunofluorescence was performed on granulosa cells (GCs) after in vitro culture. Briefly, recovered GCs after follicle puncture and GC washing were incubated in serum-free modified McCoy’s 5A medium (2.5 × 105 viable cells/well on a 8-well chamber slide (Lab-Tek® Nunc) for 48 h. Cultures were performed in a water-saturated atmosphere containing 5% CO2 in air at 38 °C. FFAR4 (green fluorescence) was immunodetected in GCs (customized FFAR4 rabbit antibody, Agro-Bio). Pre-immunized rabbit serum was used as the control with the same secondary antibody as for FFAR4 detection. Nuclei were stained with Hoechst 33,258 (blue fluorescence). The picture framed in red is a magnification of the area framed in red from the original image. Bars = 20 μm

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