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Fig. 7 | Journal of Ovarian Research

Fig. 7

From: Betacellulin regulates gap junction intercellular communication by inducing the phosphorylation of connexin 43 in human granulosa-lutein cells

Fig. 7

Effects of AG1478 and sotrastaurin on BTC-induced decreases in gap junction intercellular communication (GJIC) in SVOG cells. (A and B) SVOG cells were pretreated with DMSO, 10 µM AG1478 (A) or 10 µM sotrastaurin (B) for 40 min, and the cells were then treated with 50 ng/ml BTC for an additional 15 min. GJIC activity was determined by measuring the transfer of Lucifer yellow fluorescent dye between cells. The images were captured utilizing a fluorescence microscope (top panel). The corresponding phase contrast micrographs are shown in the bottom panel. The amount of dye transferred from one cell to its connected neighbor is dependent on the number of gap junctions that are coupled. The GJIC was estimated as an area where Lucifer yellow diffuses during a certain period away from the scrape line to the furthest extent of the dye-coupled cells. The expression or fluorescence intensities were quantified using ImageJ 1.42 software (NIH, U.S.A.). The experiments were performed in triplicate. Fluorescence intensity was calculated as fluorescence intensity of the treatment group divided by the fluorescence intensity of the control group. The image background was subtracted by ImageJ software. The scale bar represents 25 μm. Magnification, 200×. The results are expressed as the mean ± SEM of at least three independent experiments. Values marked with different letters were significantly different (P < 0.05)

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