Journal: Frontiers in Chemistry

Article Title: Synthesis, radiolabeling, and evaluation of 68Ga-labeled aminoquinoxaline derivative as a potent PFKFB3-targeted PET tracer

doi: 10.3389/fchem.2023.1158503

Figure Lengend Snippet: Structures of the representative PFKFB3 inhibitors (compounds 1–4) and compound 5.

Article Snippet: All regents used in this section were purchased from Sigma, and the Human Recombinant PFKFB3 Protein (His and GST Tag) was purchased from SinoBiological.

Techniques:

Journal: Frontiers in Chemistry

Article Title: Synthesis, radiolabeling, and evaluation of 68Ga-labeled aminoquinoxaline derivative as a potent PFKFB3-targeted PET tracer

doi: 10.3389/fchem.2023.1158503

Figure Lengend Snippet: Crystal complex of compound 4 with PFKFB3 (PDB code 6IBX) (A) and overlay between compounds 4 and 5 with PFKFB3 (B) .

Article Snippet: All regents used in this section were purchased from Sigma, and the Human Recombinant PFKFB3 Protein (His and GST Tag) was purchased from SinoBiological.

Techniques:

Journal: Frontiers in Chemistry

Article Title: Synthesis, radiolabeling, and evaluation of 68Ga-labeled aminoquinoxaline derivative as a potent PFKFB3-targeted PET tracer

doi: 10.3389/fchem.2023.1158503

Figure Lengend Snippet: Expression of PFKFB3 in the indicated human tumor tissues was analyzed using immunofluorescence staining. (A) NUGC3; (B) HCC827; (C) MKN45, (D) H1975. Scale bar, 100 µm.

Article Snippet: All regents used in this section were purchased from Sigma, and the Human Recombinant PFKFB3 Protein (His and GST Tag) was purchased from SinoBiological.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Frontiers in Chemistry

Article Title: Synthesis, radiolabeling, and evaluation of 68Ga-labeled aminoquinoxaline derivative as a potent PFKFB3-targeted PET tracer

doi: 10.3389/fchem.2023.1158503

Figure Lengend Snippet: The IC 50 of compounds 4 and 5 in biochemical activity and cellular inhibition assays.

Article Snippet: All regents used in this section were purchased from Sigma, and the Human Recombinant PFKFB3 Protein (His and GST Tag) was purchased from SinoBiological.

Techniques: Activity Assay, Inhibition

Journal: bioRxiv

Article Title: Mechanistic studies of PFKFB2 reveals a novel inhibitor of its kinase activity

doi: 10.1101/2024.12.25.630325

Figure Lengend Snippet: Development of a PFKFB kinase activity assay and screening of potential modulators. ( A ) Schematic of the coupled enzyme activity assay protocol. Purple indicates the reaction mix containing purified PFKFB isoforms. Red indicates enzymes added for the coupled assay. The ultimate product is glycerol-3-phosphate and the assay measures the disappearance of NADH at 340nm. ( B ) Plate-reader screening of candidate compounds (all at 10 μM in DMSO; wells A3 and E10 ( red boxes ) were DMSO controls). ( C ) IC50 determination of candidate inhibitor B2 (structure on inset ) for PFKFB2 ( top ) and PFKFB3 ( lower ) kinase activities. ( D ) IC50 determination of known PFKFB3 kinase inhibitor PFK158 (structure on inset ) for PFKFB2 ( top ) and PFKFB3 ( lower ) kinase activities.

Article Snippet: PFKFB2 and PFKFB3 constructs (Origene) were subcloned into GST expression vector.

Techniques: Kinase Assay, Enzyme Activity Assay, Purification

Journal: International Journal of Molecular Sciences

Article Title: PFKFB3 Inhibition Attenuates Oxaliplatin-Induced Autophagy and Enhances Its Cytotoxicity in Colon Cancer Cells

doi: 10.3390/ijms20215415

Figure Lengend Snippet: Oxa up-regulates the expression of PFKFB3. ( a ) Following treatment with Oxa for 6 h, culture media were collected and subjected to lactate assay. ( b ) Real-time PCR was performed to detect the mRNA expression of PFKFB3 following treatment with Oxa for 6 h. ( c – f ) SW480 cells were treated with Oxa ( e : 50 μM) upon to 2 h ( c : 2 h). Cell lysates were subjected to immunoblotting with the indicated antibodies, and the relatively ratios of PFKFB3 and p-Ulk1 to Actin were shown in the histogram graphs. * p < 0.05 vs. control, and ** p < 0.01 vs. control. Data represent three independent experiments.

Article Snippet: The siRNA specific for human MAP LC3β (sc-43390), PFKFB3 (sc-44011), and Ulk1 (sc-44182) were purchased from Santa Cruz Biotechnology along with the control siRNA (sc-37007).

Techniques: Expressing, Lactate Assay, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: PFKFB3 Inhibition Attenuates Oxaliplatin-Induced Autophagy and Enhances Its Cytotoxicity in Colon Cancer Cells

doi: 10.3390/ijms20215415

Figure Lengend Snippet: PFKFB3 inhibition lessens Oxa-induced autophagy. ( a , b ) Cell lysates were subjected to immunoblotting following treatment of SW480 cells with Oxa, or together with PFK-15 (6 μM; unless otherwise indicated) in the presence or absence of CQ for 2 h. ( c , d ) SW480 cells were transfected with the PFKFB3 siRNA or control siRNA (mock) for 48 h. Cell lysates were collected and subjected to immunoblotting following treatment with Oxa in the presence or absence of CQ for 2 h. All data were acquired from at least three independent experiments.

Article Snippet: The siRNA specific for human MAP LC3β (sc-43390), PFKFB3 (sc-44011), and Ulk1 (sc-44182) were purchased from Santa Cruz Biotechnology along with the control siRNA (sc-37007).

Techniques: Inhibition, Western Blot, Transfection, Control

Journal: International Journal of Molecular Sciences

Article Title: PFKFB3 Inhibition Attenuates Oxaliplatin-Induced Autophagy and Enhances Its Cytotoxicity in Colon Cancer Cells

doi: 10.3390/ijms20215415

Figure Lengend Snippet: PFKFB3 inhibition enhances the cytotoxicity of Oxa. ( a , b ) Colony growth assay was performed with Oxa (1.5 μM) in the presence or absence of PFK-15. ( c , d ) Cell viability was analyzed by MTS assay following the treatment with Oxa in the presence or absence of PFK-15 for 24 h ( c ); cell lysates from cells treated same as ( c ) were subjected to immunoblotting with indicated antibodies ( d ). ( e – h ) SW480 cells were transfected with the PFKFB3 siRNA or control siRNA for 48 h. Colony growth assay was performed with Oxa (1.5 μM) ( e ), and relative clones number to control was shown in ( f ). Cell viability was analyzed by MTS assay following the treatment with Oxa for 24 h ( g ). Cell lysates were subjected to immunoblotting with indicated antibodies following treatment with Oxa for 24 h ( h ). For histogram graph data, * p < 0.05 vs. control, and ** p < 0.01 vs. control. Similar experiments were repeated three times.

Article Snippet: The siRNA specific for human MAP LC3β (sc-43390), PFKFB3 (sc-44011), and Ulk1 (sc-44182) were purchased from Santa Cruz Biotechnology along with the control siRNA (sc-37007).

Techniques: Inhibition, Growth Assay, MTS Assay, Western Blot, Transfection, Control, Clone Assay

Journal: International Journal of Molecular Sciences

Article Title: PFKFB3 Inhibition Attenuates Oxaliplatin-Induced Autophagy and Enhances Its Cytotoxicity in Colon Cancer Cells

doi: 10.3390/ijms20215415

Figure Lengend Snippet: Primer sequences for Real-time PCR analysis.

Article Snippet: The siRNA specific for human MAP LC3β (sc-43390), PFKFB3 (sc-44011), and Ulk1 (sc-44182) were purchased from Santa Cruz Biotechnology along with the control siRNA (sc-37007).

Techniques: Real-time Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Andrographolide attenuates imbalance of gastric vascular homeostasis induced by ethanol through glycolysis pathway

doi: 10.1038/s41598-019-41417-5

Figure Lengend Snippet: Andrographolide suppresses endothelial PFKFB3 mediated glycolysis pathway. ( A ) Quantitative real time RT-PCR analysis of the expression of glycolysis related genes in HUVEC-C following andrographolide treatment. ( B ) Western blot analysis PFKFB3 expression in HUVEC-C after andrographolide treatment and the quantification data displayed in ( C ). All data presented as mean ± SEM, * P < 0.05 was considered significant. The experiments repeated three times. ( D ) IHC was performed to observe the expression of PFKFB3 in gastric wall. ( E , F ) HUVEC treated with 3-PO, a PFKFB3 inhibitor, would scratch and cell number accounting experiment performed to detect the proliferation and migration.

Article Snippet: The antibody used in this study were CD31 (Biocare; mouse, 1:200); Mac-2 (Antidodies; rabbit, 1:200); PCNA (Cell Signaling Technology; mouse, 1:1000); β-actin (Cell Signaling Technology; mouse, 1:3000); AKT (Cell Signaling Technology; rabbit, 1:2000); pAKT (Cell Signaling Technology; rabbit, 1:2000); PFKFB3 (Proteintech; rabbit, 1:1000).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Migration

Journal: Cancer Research

Article Title: Oncogene Pathway Activation in Mammary Tumors Dictates FDG-PET Uptake

doi: 10.1158/0008-5472.can-14-1235

Figure Lengend Snippet: Figure 5. Molecular correlates of FDG uptake. FDG uptake is negatively correlated with PFK-2b mRNA (A), p-PFK2 (B), and activation of AMPK (C), and positively correlated with HIF1a expression (D).

Article Snippet: Antibodies used were: Glut1 (1:5,000; kindly provided by Dr. Morris Birnbaum, University of Pennsylvania), HK2 (C4G5, 1:1,000; Cell Signaling Technology), Glut4 (1:1,000; Cell Signaling Technology), p-mTOR (p-S2448, 1:1,000; Cell Signaling Technology), p-AMPK (p-T172, 1:1,000; Cell Signaling Technology), p-S6 (p-S235/236, 1:1,000; Cell Signaling Technology), and p-PFK2 (p-S483, 1:1,000; Santa Cruz Biotechnology).

Techniques: Activation Assay, Expressing