pf 739  (MedChemExpress)

Journal: The Journal of Clinical Investigation

Article Title: Hepatic glycogen directly regulates gluconeogenesis through an AMPK/CRTC2 axis in mice

doi: 10.1172/JCI188363

Figure Lengend Snippet: ( A ) Western blots of WT and AMPKLKO hepatocytes treated with vehicle or the AMPK activator PF-739 (PF). ( B ) Western blots of WT and AMPKLKO liver lysates under refeeding (RF), short-fasting (SF), and long-fasting (LF) conditions. ( C ) Western blots of WT and PTGLKO liver lysates under SF, LF, and RF conditions. ( D ) Western blots of AML12 cell lysates. Cells were transfected with WT or S 349 D CRTC2 and treated with cycloheximide (CHX) for indicated times. ( E ) Western blots and quantification of proteins from WT and PTGLKO primary hepatocytes. Cells were treated with CHX for indicated times. RalA was used as the control. ( F ) Western blots of WT primary hepatocytes treated with different doses of glucagon or cAMP. Cells were isolated from C57BL/6J mice and pretreated with vehicle or PF for 2 hours followed by glucagon or cAMP treatment for 15 minutes. ( G ) Western blots of WT hepatocytes treated with different doses of PF. ( H ) Gene expression of Nr4a3 and Pgc1a in AML12 cells treated with cAMP. AML12 cells were transfected with vector or CRTC2 and pretreated with vehicle or PF. Data were normalized to vehicle-treated vector group shown in . ( I ) Dose-response curve for the effect of cAMP on gluconeogenic gene expression in WT and AMPKLKO primary hepatocytes. n = 3 per group. Experiments were performed at least 3 times. # * $ P < 0.05; ## ** $$ P < 0.01; ### *** $$$ P < 0.001 by 1-way ANOVA and unpaired Student’s t test. * indicates comparison with vector; # indicates comparison with vector + PF; $ indicates comparison with CRTC2.

Article Snippet: PF-739 and Ulk1 inhibitor (SBI-0206965) were purchased from MedChem Express.

Techniques: Western Blot, Transfection, Control, Isolation, Gene Expression, Plasmid Preparation, Comparison

Journal: bioRxiv

Article Title: An Enzyme-Linked Immunosorbent Assay (ELISA)-based Activity Assay for AMP-Activated Protein Kinase (AMPK)

doi: 10.1101/2024.08.20.608877

Figure Lengend Snippet: Compound C (5 μM; an AMPK inhibitor), compound 991 (250 nM; an AMPK activator), PF-739 (250 nM; an AMPK activator) or DMSO control were added into the kinase reaction (40 min reaction time) in the presence or absence of AMP. OD values from the ELISA following the kinase reactions were measured and compared, n = 4. For each treatment condition, the OD value from 0 min reaction was determined and considered as blank, and was subtracted from the OD value of the experimental condition. **** p < 0.0001 when comparing to control condition with the same AMP availability in the reaction using Student’s t-test. $ p < 0.05, $$$ p < 0.001, $$$$ p < 0.0001 when comparing to the condition with the same modulator treatment but with the presence of AMP using Student’s t-test. Data are mean ± s.e.m.

Article Snippet: AMPK inhibitor, compound C (cat# AOB2281), and AMPK activators, compound 991 (cat# AOB8150) and PF-739 (cat# AOB33584), were purchased from AOBIOUS (Gloucester, MA, USA).

Techniques: Control, Enzyme-linked Immunosorbent Assay