Fig 1: Effect of GnRH, LH, and FSH at doses of 50,100, and 150 ng/mL on omentin-1 (OMNT1) protein expression (A–C) and levels in the culture medium (A′–C′) of AP cells collected on days 10–12 of the estrous cycle of LW pigs. The protein expression was analyzed using western blot, and the results are presented as a densitometric normalized ratio relative to the β-actin abundance. The OMNT1 concentration in the culture medium was evaluated using ELISA. The results of at least four independent replicates are presented as means ± SEM for each group. Data were compared by an unpaired two-tailed Student's t test (****p < 0.0001). Representative blots are attached as Supplementary Fig. 1.
Fig 2: The effect of GnRH (100 ng/mL), FSH (100 ng/mL), LH (100 ng/mL) and INS (10 ng/mL) on visfatin protein expression (A–D) and visfatin secretion by the porcine pituitary cells (E–H) during the oestrous cycle. The visfatin protein expression was analysed by Western blot. Results are shown as representative immunoblots (each of the panels represents one blot; uncropped images of visfatin and actin immunoblots are attached in the Supplementary File 1) and bar graphs with densitometry measurement of relative visfatin protein content normalised with actin protein. Visfatin concentration in culture media was evaluated based on ELISA assay. Results are means ± S.E.M (n = 5). Values associated with bars with different superscripts are different; capital letters indicate p < 0.05. C control, GnRH gonadotrophin-releasing hormone, FSH follicle-stimulating hormone, LH luteinising hormone, INS insulin.
Fig 3: Immunofluorescence localisation of VIS and LH, FSH or ACTH in the porcine pituitary cells collected on days 10 to 12 of the oestrous cycle. Immunofluorescence labelling of VIS, DAPI (nuclear staining, NC) and LH (A–A’’’), FSH (B–B’’’), and ACTH (C–C’’’) in the porcine pituitary on days 10 to 12 of the oestrous cycle. Magnification of 40x, scale bar 20 µm. VIS visfatin, LH luteinising hormone, FSH follicle-stimulating hormone, ACTH adrenocorticotrophic hormone, NC negative control.
Fig 4: Effect of GnRH, LH, and FSH at 100 ng/mL on omentin-1 (OMNT1) protein expression (A–C) and levels in the culture medium (A′–C′) of AP cells collected on days 2–3, 10–12, 14–16, and 17–19 of the estrous cycle of Large White pigs. The protein expression was analyzed using western blot, and the results are presented as a densitometric normalized ratio relative to the β-actin abundance. The OMNT1 concentration in the culture medium was evaluated using ELISA. The results of at least four independent replicates are presented as means ± SEM for each group. Data were compared by an unpaired two-tailed Student’s t test (***p < 0.001, ****p < 0.0001). Representative blots are attached as Supplementary Fig. 1.
Fig 5: Co-localization of omentin-1 (OMNT1) in porcine tropic cells on days 10–12 of the estrous cycle in Large White pigs. The immunoreactivity of all hormones was determined by fluorescent immunohistochemistry. Left column: OMNT1 expression, visualized by Alexa Fluor 488 as green fluorescence; middle column: tropic hormone expression, visualized by Alexa Fluor 594 as red fluorescence; right column: immunofluorescent double labeling of OMNT1 and GH (A–A″), PRL (B-B”), ACTH (C–C″), TSH (D–D″), LH (E-E″), and FSH (F–F″). Arrows indicate examples of dual-labeled cells or lack of co-expression between OMNT1 and ACTH (C–C′). Scale bar: 20 μm. Section thickness 5 μm.
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