β 2 ar  (Bioss)

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) ADRB2 expression in TCGA cancer tissues compared with corresponding TCGA and GTEx normal tissues. (B) ADRB2 expression in 7 types of cancer based on TCGA cancer and normal sample data. (C) Association between overall survival and ADRB2 expression in LUAD. (D) Immunoblot image of ADRB2 in LUAD and LUSC cell lines. ADRB2, adrenoceptor β2; GTEx, Genotype-Tissue Expression; TCGA, The Cancer Genome Atlas. * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, Western Blot

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Percentages of wt, del and amp in LUAD. (B) Correlation between copy number variations and ADRB2 mRNA expression in LUAD samples. (C) Correlation between amp and ADRB2 mRNA expression in LUAD samples. (D) Correlation between dels and ADRB2 mRNA expression in LUAD samples. (E) Correlation between DNA methylation and ADRB2 mRNA in LUAD samples. (F) miRNA-ADRB2 regulatory network generated using Cytoscape software. (G) Examination of miR-424-5p expression in LUAD and normal samples conducted using the StarBase database. (H) Result of dual luciferase reporter gene experiment. ADRB2, adrenoceptor β2; amp, amplification; del, deletion; LUAD, lung adenocarcinoma; miR/miRNA, microRNA; wt, wild-type.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, DNA Methylation Assay, Generated, Software, Luciferase, Amplification

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Various immune cells infiltrate LUAD levels in case of different copy numbers of ADRB2. (B) Correlation of ADRB2 expression with infiltration levels of immune cells in LUAD. (C) and (D) ADRB2 expression specifically within these immune cell types at the single-cell level. (E) Chemokine receptor genes. (F) Chemokine genes. (G) Immune activation genes. (H) Immunosuppressive genes. ADRB2, adrenoceptor β2; LUAD, lung adenocarcinoma. Correlation between adrenoceptor β2 and tumor immunity-related genes.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, Single Cell, Activation Assay

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Validation of stable ADRB2 gene overexpression in LUAD Cell lines. (B) Comparison of tumor volume between control and ADRB2-overexpressing groups. (C) Visualization of tumorigenesis via small animal live imaging. (D) Comparison of tumor size between the control and ADRB2-overexpressing groups in C57BL/6 mice. Tumors in mice with ADRB2-overexpressing LUAD cells were significantly smaller, indicating the suppressive effect of ADRB2 on tumor growth. (E) Flow cytometry analysis of CD4 + T cell infiltration in tumor tissues from C57BL/6 mice (CD45 + CD11b - CD45R - CD3 + CD4 + ). (F) Flow cytometry analysis of dendritic cell infiltration in tumor tissues from C57BL/6 mice (CD45 + CD11c + MHC II + ). ** P < 0.01, *** P < 0.01. ADRB2, adrenoceptor β2.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Biomarker Discovery, Over Expression, Comparison, Control, Imaging, Flow Cytometry

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Correlation of ADRB2 and PD-1 expression in LUAD. (B) Correlation of ADRB2 and PD-L1 expression in LUAD. (C) Correlation of ADRB2 and CTLA-4 expression in LUAD. (D) The correlation of ADRB2 and PD-1 expression in LUAD was verified using the GEPIA database. (E) The correlation of ADRB2 and PD-L1 expression in LUAD was verified using the GEPIA database. (F) The correlation of ADRB2 and CTLA-4 expression in LUAD was verified using the GEPIA database. ADRB2, adrenoceptor β2; CTLA-4, cytotoxic T-lymphocyte associated protein 4; GEPIA, Gene Expression Profiling Interactive Analysis; LUAD, lung adenocarcinoma; PD-1, programmed cell death protein 1; PD-L1, programmed death-ligand 1.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, Gene Expression

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: Data from the GDSC shows the association between ADRB2 expression and drug sensitivity.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: ADRB2, adrenoceptor β2; miR, microRNA.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques:

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) ADRB2 expression in TCGA cancer tissues compared with corresponding TCGA and GTEx normal tissues. (B) ADRB2 expression in 7 types of cancer based on TCGA cancer and normal sample data. (C) Association between overall survival and ADRB2 expression in LUAD. (D) Immunoblot image of ADRB2 in LUAD and LUSC cell lines. ADRB2, adrenoceptor β2; GTEx, Genotype-Tissue Expression; TCGA, The Cancer Genome Atlas. * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, Western Blot

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Percentages of wt, del and amp in LUAD. (B) Correlation between copy number variations and ADRB2 mRNA expression in LUAD samples. (C) Correlation between amp and ADRB2 mRNA expression in LUAD samples. (D) Correlation between dels and ADRB2 mRNA expression in LUAD samples. (E) Correlation between DNA methylation and ADRB2 mRNA in LUAD samples. (F) miRNA-ADRB2 regulatory network generated using Cytoscape software. (G) Examination of miR-424-5p expression in LUAD and normal samples conducted using the StarBase database. (H) Result of dual luciferase reporter gene experiment. ADRB2, adrenoceptor β2; amp, amplification; del, deletion; LUAD, lung adenocarcinoma; miR/miRNA, microRNA; wt, wild-type.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, DNA Methylation Assay, Generated, Software, Luciferase, Amplification

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Various immune cells infiltrate LUAD levels in case of different copy numbers of ADRB2. (B) Correlation of ADRB2 expression with infiltration levels of immune cells in LUAD. (C) and (D) ADRB2 expression specifically within these immune cell types at the single-cell level. (E) Chemokine receptor genes. (F) Chemokine genes. (G) Immune activation genes. (H) Immunosuppressive genes. ADRB2, adrenoceptor β2; LUAD, lung adenocarcinoma. Correlation between adrenoceptor β2 and tumor immunity-related genes.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, Single Cell, Activation Assay

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Validation of stable ADRB2 gene overexpression in LUAD Cell lines. (B) Comparison of tumor volume between control and ADRB2-overexpressing groups. (C) Visualization of tumorigenesis via small animal live imaging. (D) Comparison of tumor size between the control and ADRB2-overexpressing groups in C57BL/6 mice. Tumors in mice with ADRB2-overexpressing LUAD cells were significantly smaller, indicating the suppressive effect of ADRB2 on tumor growth. (E) Flow cytometry analysis of CD4 + T cell infiltration in tumor tissues from C57BL/6 mice (CD45 + CD11b - CD45R - CD3 + CD4 + ). (F) Flow cytometry analysis of dendritic cell infiltration in tumor tissues from C57BL/6 mice (CD45 + CD11c + MHC II + ). ** P < 0.01, *** P < 0.01. ADRB2, adrenoceptor β2.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Biomarker Discovery, Over Expression, Comparison, Control, Imaging, Flow Cytometry

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: (A) Correlation of ADRB2 and PD-1 expression in LUAD. (B) Correlation of ADRB2 and PD-L1 expression in LUAD. (C) Correlation of ADRB2 and CTLA-4 expression in LUAD. (D) The correlation of ADRB2 and PD-1 expression in LUAD was verified using the GEPIA database. (E) The correlation of ADRB2 and PD-L1 expression in LUAD was verified using the GEPIA database. (F) The correlation of ADRB2 and CTLA-4 expression in LUAD was verified using the GEPIA database. ADRB2, adrenoceptor β2; CTLA-4, cytotoxic T-lymphocyte associated protein 4; GEPIA, Gene Expression Profiling Interactive Analysis; LUAD, lung adenocarcinoma; PD-1, programmed cell death protein 1; PD-L1, programmed death-ligand 1.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing, Gene Expression

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: Data from the GDSC shows the association between ADRB2 expression and drug sensitivity.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques: Expressing

Journal: PLOS One

Article Title: C1RL-AS1/microRNA-424-5p/adrenoceptor β2 axis: A novel regulatory mechanism in lung adenocarcinoma associated with immune infiltration and prognosis

doi: 10.1371/journal.pone.0343805

Figure Lengend Snippet: ADRB2, adrenoceptor β2; miR, microRNA.

Article Snippet: ADRB2 was determined with the primary anti-ADRB2 antibody (29864–1-AP, Wuhan Proteintech Group, Inc., Ltd) and secondary antibody (SA00001–2, Wuhan Proteintech Group, Inc., Ltd).

Techniques:

Journal: Nature Communications

Article Title: An artificial cell capable of signal transduction mediated by ADRB2 for the regulation of glycogenolysis

doi: 10.1038/s41467-026-68503-3

Figure Lengend Snippet: a The signal transduction from extracellular ISO into intracellular cAMP and subsequent regulation of glycogenolytic metabolic pathway inside an artificial cell; ISO: isoproterenol; ADRB2: β2-adrenergic receptor; Gsα: Gs subunit α; ADCY5: adenylate cyclase V; PKA: Protein kinase A; PhK: Phosphorylase kinase; PYGM: Glycogen phosphorylase; PGM: Phosphoglucomutase; G6PDH: Glucose-6-phosphate dehydrogenase; GDP: Guanosine diphosphate; GTP: Guanosine triphosphate; ATP: Adenosine triphosphate; cAMP: Cyclic adenosine monophosphate; G-1-P: Glucose 1-phosphate; G-6-P: Glucose-6-phosphate; 6-PGL: 6-Phosphoglucono-lactone; NADP⁺(H): Nicotinamide adenine dinucleotide phosphate. Created in BioRender. Liu, Y. (2025) https://BioRender.com/689u8j4 . b Chemical reaction equations involved in the signal transduction and glycogenolytic metabolism; ADCY5: Adenylate cyclase V; p-PYGM: Phosphorylated glycogen phosphorylase.

Article Snippet: Rabbit anti-PhKA2 polyclonal antibody (No. 24658-1-AP), rabbit anti-PYGM-Specific polyclonal antibody (No. 19716-1-AP) and ADRB2 polyclonal antibody (No. 29864-1-AP) were purchased from Proteintech (China).

Techniques: Transduction

Journal: Nature Communications

Article Title: An artificial cell capable of signal transduction mediated by ADRB2 for the regulation of glycogenolysis

doi: 10.1038/s41467-026-68503-3

Figure Lengend Snippet: a Scheme of the signal transduction pathway from ISO to cAMP mediated by ADRB2. SDS-PAGE images of purified ADRB2 ( b ), Gsα ( c ) and ADCY5 ( d ) from Homo Sapiens . n = 3 independent replicates. e The size exclusion chromatography of ADRB2, Gsα, and ADRB2-Gsα complex. f The effect of pH on the ADCY5 activity by measuring the production of cAMP at 30 °C, 10.0 μM ISO and 2.0 mM Mg 2+ . g The effect of temperature on the ADCY5 activity by measuring the production of cAMP at pH 8, 10.0 μM ISO and 2.0 mM Mg 2+ . The data were expressed as mean ± SD, n = 3 independent replicates. h The Mg 2+ concentration effect on the ADCY5 activity by measuring the production of cAMP at 37 °C, pH 8 and 10.0 μM ISO. The data were expressed as mean ± SD, n = 3 independent replicates. i Initial reaction velocities of cAMP production catalyzed by ADCY5 as a function of ATP concentration from 0 to 14.0 mM in the solution (10.0 μM ISO, 2.0 mM Mg 2+ , 6.0 μg/mL ADRB2-Gsα complex, 6.0 μg/mL ADCY5, pH 8) at 37 °C. The data were expressed as mean ± SD, n = 3 independent replicates. j The cAMP production as a function of ADRB2-Gsα/ADCY5 molar ratios with ADCY5 fixed at 10 nM and ADRB2-Gsα varied from 1.1 to 21.7 nM in a solution (10.0 μM ISO, 2.0 mM Mg 2+ , pH 8) at 37 °C. The data were expressed as mean ± SD, n = 3 independent replicates. **** P < 0.0001, ns P > 0.05, two-sided, multiple comparisons by One-way ANOVA test. k ISO dose-dependent cAMP production as a function of time in a solution (17.4 nM ADRB2-Gsα, 10.0 nM ADCY5, 10 −5 −10 3 μM ISO, 2.0 mM Mg 2+ , pH 8) at 37 °C. l The corresponding cAMP production at 25 min of ( k ). The data were expressed as mean ± SD, n = 3 independent replicates. **** P < 0.0001, ns P > 0.05, two-sided, multiple comparisons by One-way ANOVA test. P < 0.05 was considered statistically significant. Source data are provided as a Source Data file.

Article Snippet: Rabbit anti-PhKA2 polyclonal antibody (No. 24658-1-AP), rabbit anti-PYGM-Specific polyclonal antibody (No. 19716-1-AP) and ADRB2 polyclonal antibody (No. 29864-1-AP) were purchased from Proteintech (China).

Techniques: Transduction, SDS Page, Purification, Size-exclusion Chromatography, Activity Assay, Concentration Assay

Journal: Nature Communications

Article Title: An artificial cell capable of signal transduction mediated by ADRB2 for the regulation of glycogenolysis

doi: 10.1038/s41467-026-68503-3

Figure Lengend Snippet: a The representative fluorescence images of Cy5-ADRB2-GUV (with NBD-PE in the bilayer) taken with red channel (left image), green channel (middle image), and their merged image (right image) at ADRB2 and lipids molar ratio of 10 3 :10 6 . The scale bar is 10.0 μm. b The percentage of ADRB2-GUVs with varied ADRB2 to lipids molar ratios ranging from 1:10 6 to 10 4 :10 6 . The data were expressed as mean ± SD, n = 3 independent replicates. **** P < 0.0001 1:10 6 vs. 10:10 6 , **** P < 0.0001 10:10 6 vs. 10 2 :10 6 , *** P = 0.0007 10 2 :10 6 vs. 10 3 :10 6 , ns P > 0.05 10 3 :10 6 vs. 10 4 :10 6 , two-sided, multiple comparisons by One-way ANOVA test. c The percentage of ADRB2 with correct orientation on GUVs determined by WB (top image) and band intensities (bottom graph). The data were expressed as mean ± SD, n = 3 independent replicates. d Schematic diagram of the Gsα detection mechanism using BODIPY TR GTP γ S. Created in BioRender. Liu, Y. (2025) https://BioRender.com/g8zgjzy . e The representative time-series images of GUVs stimulated by 1.0 μM ISO for the binding events of BODIPY TR GTP γ S to Gsα from 30 s to 15 min. The scale bar is 10.0 μm. n = 3 independent replicates. f The corresponding fluorescence intensity of GUV in ( e ) (red curve) and the fluorescence intensity of GUV with no ISO stimulation (blue curve). n = 3 independent replicates. g The representative fluorescence images of FITC labeled ADCY5-GUV (with TR-DHPE in the bilayer) taken with green channel (left image), red channel (middle image), and their merged image (right image) at ADCY5 and lipids molar ratio of 10 3 :10 6 . The scale bar is 10.0 μm. n = 3 independent replicates. h The flow cytometry histogram of FITC-ADCY5-GUVs with varied ADCY5 to lipids molar ratios ranging from 10 −1 :10 6 to 10 3 :10 6 . n = 3 independent replicates. i The percentage of ADCY5-GUVs with varied ADCY5 to lipids molar ratios ranging from 10 −1 :10 6 to 10 3 :10 6 . The data were expressed as mean ± SD, n = 3 independent replicates. **** P < 0.0001 10 −1 :10 6 vs. 1:10 6 , **** P < 0.0001 1:10 6 vs. 10:10 6 , **** P < 0.0001 10:10 6 vs. 10 2 :10 6 , **** P < 0.0001 10 2 :10 6 vs. 10 3 :10 6 , two-sided, multiple comparisons by One-way ANOVA test. P < 0.05 was considered statistically significant. Source data are provided as a Source Data file.

Article Snippet: Rabbit anti-PhKA2 polyclonal antibody (No. 24658-1-AP), rabbit anti-PYGM-Specific polyclonal antibody (No. 19716-1-AP) and ADRB2 polyclonal antibody (No. 29864-1-AP) were purchased from Proteintech (China).

Techniques: Fluorescence, Binding Assay, Labeling, Flow Cytometry

Journal: Nature Communications

Article Title: An artificial cell capable of signal transduction mediated by ADRB2 for the regulation of glycogenolysis

doi: 10.1038/s41467-026-68503-3

Figure Lengend Snippet: a Schematic illustration of the signal transduction of an artificial cell from extracellular ISO to intracellular cAMP. Created in BioRender. Liu, Y. (2025) https://BioRender.com/ik5i982 . b Fluorescence images of GUVs containing Cy5-ADRB2-Gsα complexes and FITC-ADCY5 taken by red channel (left image), green channel (middle image), and their merged image (right image). The scale bar is 10.0 μm. n = 3 independent replicates. c The representative time-series images of artificial cells stimulated by 1.0 μM ISO for the intracellular cAMP production from 0 to 20 min, with top row images taken by the blue channel and bottom row images taken by the green channel. The scale bar is 10.0 μm. n = 3 independent replicates. d The corresponding FRET ratios of Epac1-cAMP in the artificial cell in ( c ). The data were expressed as mean ± SD, n = 3 independent replicates. e The cAMP production in artificial cells with the addition of 1.0 μM ISO as a function of time from 0 to 30 min using fluorescence spectroscopy. n = 3 independent replicates. f The cAMP production in artificial cells with the addition of varied ISO concentration (0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 4.0, 6.0, 8.0, 10.0 μM) at 30 min. n = 3 independent replicates. g The signal amplification folds of artificial cells from extracellular ISO to intracellular cAMP as a function of ISO concentration (0.4 to 10.0 μM) at 30 min. n = 3 independent replicates. h The representative time-series images of Epac1-cAMP in artificial cells with sequential antagonist alprenolol treatment for 10 min from 10 to 20 min, and competitive 10.0 μM ISO activation for 10 min from 20 to 30 min, with top row images taken by blue channel and bottom row images taken by green channel. The scale bar is 10.0 μm. n = 3 independent replicates. i The corresponding Fret ratio of Epac1-cAMP in the artificial cell in ( h ). The data were expressed as mean ± SD, n = 3 independent replicates. j The representative time-series images of Epac1-cAMP in artificial cells with sequential inverse agonist carazolol treatment for 10 min from 10 to 20 min, and 10.0 μM ISO activation for 10 min from 20 to 30 min, with top row images taken by blue channel and bottom row images taken by green channel. The scale bar is 10.0 μm. n = 3 independent replicates. k The corresponding FRET ratio Epac1-cAMP in the artificial cell in ( j ). The data were expressed as mean ± SD, n = 3 independent replicates. l FRET ratio of Epac1-cAMP in the artificial cell measured by fluorescence spectroscopy as a function of time with the addition of 1.0 μM alprenolol antagonism at 10 min and the addition of 10.0 μM ISO at 20 min (blue curve), with the addition of 1.0 μM carazolol inverse agonism at 10 min and the addition of 10.0 μM ISO at 20 min (green curve). The data were expressed as mean ± SD, n = 3 independent replicates. Source data are provided as a Source Data file.

Article Snippet: Rabbit anti-PhKA2 polyclonal antibody (No. 24658-1-AP), rabbit anti-PYGM-Specific polyclonal antibody (No. 19716-1-AP) and ADRB2 polyclonal antibody (No. 29864-1-AP) were purchased from Proteintech (China).

Techniques: Transduction, Fluorescence, Spectroscopy, Concentration Assay, Amplification, Activation Assay