Journal: American Journal of Physiology - Cell Physiology

Article Title: Glycolytic enzyme PFKFB3 regulates sphingosine 1-phosphate receptor 1 in proangiogenic glomerular endothelial cells under diabetic condition

doi: 10.1152/ajpcell.00261.2023

Figure Lengend Snippet: Primer information

Article Snippet: Anti-angiopoietin2 (Cat. No. NB110-85467, 1:1,000 for WB, 1:200 for IHC, RRID:AB_1199462), and phosphodiesterase 3A (PDE3A, Cat. No. NBP1-46182, 1:1,000 WB, RRID:AB_10009414) antibodies were purchased from Novus (Centennial, CO).

Techniques:

Journal: American Journal of Physiology - Cell Physiology

Article Title: Glycolytic enzyme PFKFB3 regulates sphingosine 1-phosphate receptor 1 in proangiogenic glomerular endothelial cells under diabetic condition

doi: 10.1152/ajpcell.00261.2023

Figure Lengend Snippet: A : microRNA-590-3p expressions in control and proangiogenic glomerular endothelial cells (GECs) incubated without (Ctrl) or with high glucose (HG) or mannitol (Mann), the osmatic control of high glucose. Messenger RNA expressions of the rate-limiting enzymes Glut1 ( B ), Glut4 ( C ), hexokinase 2 (HK2, D ), phosphofructokinase/fructose bisphosphatase 3 (PFKFB3, E ), pyruvate kinase enzyme M2 (PKM2, F ) in glycolysis in GECs in the presence (the proangiogenic cells) or the absence of the sprouting regimen. Representative Western blots ( G ) and densitometric quantification of PFKFB3 in proangiogenic GECs ( H ), transfected without (ctrl) or with the negative control miRNA (NC) or miRNA-590-3p mimic (mimic), after 6-h stimulation; n = 6, * P < 0.05 compared with the cells under control (without stimulation), # P < 0.05 compared with control cells stimulated with high glucose and sprouting regimen using one-way ANOVA. I : representative Western blots of nitric oxide synthase, endothelial (eNOS), protein kinase A (PKA), phosphodiesterase 3A (PDE3A), and muscle-associated glycogen phosphorylase (PYGM) protein. Densitometric quantification of total eNOS ( J ), phosphorylated PKA ( K ), PDE3A ( L ), and PYGM ( M ) protein in proangiogenic GECs. n = 5 or 6, # P < 0.05 compared with human renal glomerular endothelial cell (HGEC) stimulated with high glucose and sprouting regimen using one-way ANOVA. N : glycated rate measured by seahorse influx analyses in proangiogenic GECs, transfected without or with miRNA-590-3p mimic. n = 6. O : regulation of PFKFB3 by miRNA-590-3p in Luciferase assay. n = 4. * P < 0.05 compared with the cells under control using one-way ANOVA.

Article Snippet: Anti-angiopoietin2 (Cat. No. NB110-85467, 1:1,000 for WB, 1:200 for IHC, RRID:AB_1199462), and phosphodiesterase 3A (PDE3A, Cat. No. NBP1-46182, 1:1,000 WB, RRID:AB_10009414) antibodies were purchased from Novus (Centennial, CO).

Techniques: Control, Incubation, Western Blot, Transfection, Negative Control, Luciferase