ampk antagonist compound c  (ApexBio)

Journal: Cells

Article Title: APN Expression in Serum and Corpus Luteum: Regulation of Luteal Steroidogenesis Is Possibly Dependent on the AdipoR2/AMPK Pathway in Goats

doi: 10.3390/cells12101393

Figure Lengend Snippet: APN activates the AMPK signal in goat luteal steroidogenic cells. ( A ) Luteal tissue was paraffin-embedded and processed for mIHC. P-AMPK (dark red), AMPK (olive) and DAPI staining (blue) for nucleus. NC, negative control; SLC, small luteal cells; LLC, large luteal cells. Scale bar = 20 µm. ( B ) WB for P-AMPK, AMPK and STAR of cyclic CLs. ( C ) WB for P-AMPK in mid-cycle and pregnant CLs. M, mid-cycle; P, pregnancy. ( D ) Cells were treated with 1 μg/mL APN (left) or 25 μM AdipoRon (right) for 0, 0.5, 1 and 2 h to detect P-AMPK by WB. ( E ) Cells were treated with 1 μg/mL APN or 25 μM AdipoRon for 1 h to detect whole and cytoplasmic APPL1 and LKB1 by WB. Days 0, 4, 11 and 17 represent the days in the estrous cycle, with day 0 as the onset of estrus. Data are shown as means ± SEM. A Student’s t -test was used for the comparison of two datasets. A one-way analysis of variance was followed by a Tukey’s multiple comparisons test. Statistical significance is indicated with asterisks *, ** and ***, reflecting p values of <0.05, p < 0.01 and p < 0.001, respectively.

Article Snippet: Cells were treated with APN (Sangon, Shanghai, China) [ ], AdipoRon (MedChemExpress, Monmouth Junction, NJ, USA) [ ] or AMPK antagonist (Compound C, Apexbio, Houston, TX, USA) [ , ].

Techniques: Staining, Negative Control, Comparison

Journal: Cells

Article Title: APN Expression in Serum and Corpus Luteum: Regulation of Luteal Steroidogenesis Is Possibly Dependent on the AdipoR2/AMPK Pathway in Goats

doi: 10.3390/cells12101393

Figure Lengend Snippet: APN/AdipoRon modulates steroidogenesis through the AMPK signal in goat luteal cells. ( A ) Luteal cells were pretreated with Compound C (10 μM, 1 h) prior to treatment with AdipoRon (25 μM, 1 h), and cell lysates were processed to detect P-AMPK by WB. Luteal cells were pretreated with Compound C (10 μM, 1 h) prior to treatment with AdipoRon (25 μM, 24 h), media were collected to detect the P4 concentration by RIA ( B , upper) and cell lysates were processed to detect STAR, CYP11A1 and HSD3B by WB ( C ). ( D ) Luteal cells were pretreated with SiAMPK for 48 h, then cells were treated with 1 μg/mL APN for 24 h to detect AMPK, STAR, CYP11A1 and HSD3B by WB, and media were collected to detect the P4 concentration by RIA ( B , lower). SiAMPK, SiRNA for AMPK; scrambled non-targeting RNA was transfected into cells as a negative control. Data are expressed as means ± SEM and the statistical difference was analyzed by a one-way ANOVA followed by a Tukey’s multiple comparisons test. * and ** reflect p values of <0.05 and p < 0.01, respectively.

Article Snippet: Cells were treated with APN (Sangon, Shanghai, China) [ ], AdipoRon (MedChemExpress, Monmouth Junction, NJ, USA) [ ] or AMPK antagonist (Compound C, Apexbio, Houston, TX, USA) [ , ].

Techniques: Concentration Assay, Transfection, Negative Control