Fig. 2

A Representative image of western blot analysis of pERK 1/2 and total ERK 1/2 expression in GCs (n = 20) were measured under various conditions: untreated (Ctrl), treated with BCA 10 µM for 24-h, and co-treated with BCA 10 µM plus either PRO 1 mM (BCA + PRO), SBA 30 µM (BCA + SBA), or CITRO 150 µM (BCA + CITRO) for 24-h. Antagonists were preincubated for 1 h before the addition of BCA. Equal protein loading of the preparations was verified using Ponceau staining. The image is representative of three independent experiments. B-D Computer-assisted semi-quantitative analysis of the overall relative intensity of the bands. The intensity was measured (pixel/mm2) and then normalized relative to Ponceau staining. Values are expressed as fold change as respect to the control. Different letters indicate statistically significant differences (p ≤ 0.05). E Intracellular cAMP concentration in GCs. Results (expressed as mean ± SD) are expressed as a percentage relative to untreated cells, which are set at 100%. Experiments were each performed in duplicate. F Intracellular Calcium levels (mM) in GCs according to different treatments, measured in duplicate. Statistical analyses were conducted using GraphPad Prism 9, employing a one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test to assess significant differences. Different letters indicate statistically significant differences (p ≤ 0.05)