Figure 4

Western-immunodetection of RXFP1 and RXFP2 protein. The figure shows representative gel electrophoreses of at least three independent replicates. A: Validation of anti-human RXFP1 and RXFP2 in porcine ovarian granulosa cells (GC) and corpora lutea (CL). After gel electrophoreses, resolved proteins were transferred to PVDF membranes and before exposure to antibodies, membranes were pre-incubated (GC+ and CL+) or not (GC- and CL-) with RXFP1 or RXFP2 ligands (relaxin and INSL3, respectively). B: Membranes containing total proteins isolated from porcine granulosa cells (GC), immature (ICC) and mature (MCC) cumulus cells, immature (GV) and mature (MII) oocyte, and spermatozoa (SP) were incubated with anti-human RXFP1 or RXFP2 antibodies. Arrows indicate corresponding RXFP1 (≈82 kDa) and RXFP2 (≈62 kDa) bands. Band quantifications (using Image J) are presented in associated figures with statistic analyses of band intensities. Asterisks and letters indicate significant differences within the same (*) or between (a,b,c,d) cell types (P < 0.05; Student's t-Test).