Figure 2

Expression and characterization of genes belonging to SFKs and their functional activation in the monkey CL throughout the luteal phase. (A) Diagrammatic representation of various signalling cascades and molecular players involved downstream of LH/CGR activation regulating various genes associated with steroidogenesis. Based on the review of literature the canonical pathway Gs/cAMP/protein kinase A (PKA)/pCREB and alternate signalling pathways like p38 MAPK or Erk have been illustrated in the diagram. Furthermore, recent studies describing involvement of SFKs and cAMP-PDE in modulation of hCG responsiveness and transactivation of tyrosine kinase receptors in mouse Leydig tumor cells are also represented for their possible similar role in the monkey luteal cells. (B) Semi-quantitative RT-PCR expression of genes belonging to SFKs (Fyn, Yes and Src) in the monkey CL obtained from different stages of the luteal phase. Each bar represents mean ± SEM values (n = 3 CL/stage). L-19 mRNA was used as internal control and relative expression was calculated following densitometric analysis. Bars with different letters indicate statistical significance (p < 0.05). (C) Levels of Src family of kinase protein in the monkey CL throughout the luteal phase. Immunoblot analysis was performed to determine functional activation of Src protein i.e., protein levels of active pSrc (Y-416), inactive pSrc (Y-527) and total Src in the monkey CL collected during different stages of the luteal phase. Anti-β-actin antibody (the protein loading control) probed blot is presented to indicate equal amount of protein being loaded in each lane. Densitometric analysis of immunoblots was determined and is indicated as mean ± SEM of relative amount of pSrc (Y-416/Y-527) expressed as intensity of total Src/β-actin bands for each stage of luteal phase (n = 3 CL/stage). Bars with different letters indicate statistical significance (p < 0.05).