Journal: PLoS ONE

Article Title: Monocyte Gene Expression Signature of Patients with Early Onset Coronary Artery Disease

doi: 10.1371/journal.pone.0032166

Figure Lengend Snippet: The x-axis the average change in expression following the use of statins and aspirin depicted as 2expΔΔCT. Values are given as mean + SD. A relative fold change of ≤1 or −1 indicates no effect. For ADRB2 and ABCG1 the changes were significant; p = 0.04 and p<0.001 respectively.

Article Snippet: Differential expression was confirmed by qPCR for ABCA1 (Hs02565169_s1); ABCG1 (Hs00245154_m1); RGS1 (Hs00175260_m1); ADRB2 (Hs00240532_s1); FOLR3 (Hs00357145_g1) and GSTM1 (Hs02341469_m1) by qPCR.

Techniques: Expressing

Journal: The FASEB Journal

Article Title: α-Ketoglutarate prevents skeletal muscle protein degradation and muscle atrophy through PHD3/ADRB2 pathway

doi: 10.1096/fj.201700670R

Figure Lengend Snippet: PCR primer sequences and amplification parameters

Article Snippet: C2C12 cells were ncubated overnight in rabbit anti-PHD3 (bs-0532R; Bioss Antibodies) and rabbit anti-ADRB2 (bs-0947R; Bioss Antibodies) at 4°C.

Techniques: Amplification, Sequencing

Journal: The FASEB Journal

Article Title: α-Ketoglutarate prevents skeletal muscle protein degradation and muscle atrophy through PHD3/ADRB2 pathway

doi: 10.1096/fj.201700670R

Figure Lengend Snippet: ADRB2 is target of PHD3 in response to AKG. A) C2C12 cells were induced differentiation for 4 d, and anti-PHD3 was used to pull down PHD3 binding protein. Precipitated samples were used for SDS-PAGE, and then strips were stained by Coomassie Brilliant Blue. B) C2C12 myotubes were treated by 2 mM AKG for 48 h, and anti-PHD3 and anti-ADRB2 were used to precipitate PHD3 and ADRB2, respectively. Precipitated samples was subjected to immunoblotting of ADRB2 or PHD3 (n = 3 group). C) C2C12 myotube samples from control or 2 mM AKG–treated group were pulled down by anti-ubiquitin antibody and subjected to immunoblotting of ADRB2 (n = 3/group). D) Immunoblots and quantification of ADRB2 in normal or PHD3-overexpressing C2C12 myotubes treated with 2 mM AKG for 48 h (n = 6/group). E, F) Representative images (E) and quantification (F) of ADRB2 in C2C12 myotubes treated with 0 or 2 mM AKG for 48 h (n = 6/group). G) Immunoblots and quantification of ADRB2 in C2C12 myotubes treated with 0 or 2 mM AKG for 48 h (n = 6/group). H) cAMP levels in C2C12 myotubes treated with 0 or 2 mM AKG for 48 h (n = 6/group). I–K) Immunoblots (I) and quantification (J, K) of p-PKA (J), PKA, p-CREB (K), and CREB (K) in C2C12 myotubes treated with 0 or 2 mM AKG for 48 h (n = 6/group). Data are presented as means ± sem and were analyzed by 1-way ANOVA followed by post hoc Bonferroni tests for panel D, and nonpaired Student’s t test for all others. β-Actin served as housekeeping gene. *P < 0.05 compared to control.

Article Snippet: C2C12 cells were ncubated overnight in rabbit anti-PHD3 (bs-0532R; Bioss Antibodies) and rabbit anti-ADRB2 (bs-0947R; Bioss Antibodies) at 4°C.

Techniques: Binding Assay, SDS Page, Staining, Western Blot

Journal: The FASEB Journal

Article Title: α-Ketoglutarate prevents skeletal muscle protein degradation and muscle atrophy through PHD3/ADRB2 pathway

doi: 10.1096/fj.201700670R

Figure Lengend Snippet: Pharmacologic inhibition of ADRB2 blocked inhibitory effects of AKG on skeletal muscle atrophy and protein degradation. A–C) Immunoblots (A) and quantification (B, C) of p-PKA and PKA (B) and p-CREB and CREB (C) in C2C12 myotubes treated with vehicle, 2 mM AKG, 10 μM ICI, or 2 mM AKG + 10 μM ICI for 48 h (n = 6/group). D, E) Representative images (D) and quantification (E) of long-life protein Click-It AHA in C2C12 myotubes treated with vehicle, 2 mM AKG, 10 μM ICI, or 2 mM AKG + 10 μM ICI for 48 h (n = 6/group). F, G) Representative images (F) and quantification (G) of fiber diameter of C2C12 myotubes treated with vehicle, 2 mM AKG, 10 μM ICI, or 2 mM AKG + 10 μM ICI for 48 h (n = 6/group). H–K) Immunoblots (H) and quantification (I–K) of pFoxO1 and FoxO1 (I), MuRF1 (J), and MAFbx (K) in C2C12 myotubes treated with vehicle, 2 mM AKG, 10 μM ICI, or 2 mM AKG + 10 μM ICI for 48 h (n = 6/group). L, M) Representative images (L) and quantification (M) of propidium iodide (PI, red)-positive MuRF1 (green) cells (yellow nucleus indicated by white arrows) in gastrocnemius muscle from male C57BL/6J mice 3 h after intraperitoneal injection with vehicle, 1 g/kg AKG, 5 μg/kg ICI, or 1 g/kg AKG + 5 μg/kg ICI (n = 6). N) 3-MeHis in gastrocnemius muscle from male C57BL/6J mice 3 h after intraperitoneal injection with vehicle, 1 g/kg AKG, 5 μg/kg ICI, or 1 g/kg AKG + 5 μg/kg ICI (n = 6). O–R) Immunoblots (O) and quantification (P–R) of pFoxO1 and FoxO1 (P), MuRF1 (Q), and MAFbx (R) in gastrocnemius muscle from male C57BL/6J mice 3 h after injected intraperitoneally with vehicle, 1 g/kg AKG, 5 μg/kg ICI, or 1 g/kg AKG + 5 μg/kg ICI (n = 6). Data are presented as means ± sem and were analyzed by 1-way ANOVA, followed by post hoc Bonferroni tests. β-Actin served as housekeeping gene. *P < 0.05 compared to control.

Article Snippet: C2C12 cells were ncubated overnight in rabbit anti-PHD3 (bs-0532R; Bioss Antibodies) and rabbit anti-ADRB2 (bs-0947R; Bioss Antibodies) at 4°C.

Techniques: Inhibition, Western Blot, Injection

Journal: The FASEB Journal

Article Title: α-Ketoglutarate prevents skeletal muscle protein degradation and muscle atrophy through PHD3/ADRB2 pathway

doi: 10.1096/fj.201700670R

Figure Lengend Snippet: ADRB2 knockdown abolished inhibitory effects of AKG on muscle atrophy and protein degradation. A) mRNA expression of ADRB2 in gastrocnemius muscle from male C57BL/6J mice injected intramuscularly with LV-shScrambled or LV-shADRB2. B–D) Immunoblots (B) and quantification (C, D) of p-PKA and PKA (C) and p-CREB and CREB (D) in gastrocnemius muscle from male C57BL/6J mice receiving LV-shScrambled, LV-shScrambled + AKG, LV-shADRB2, or LV-shADRB2 +AKG. E–J) Gastrocnemius weight (E), gastrocnemius muscle fiber size (F, G), muscle grip (H), high-speed running time (I), and slow-speed running time (J) of male C57BL/6J mice receiving LV-shScrambled, LV-shScrambled + AKG, LV-shADRB2, or LV-shADRB2 + AKG. K–N) Immunoblots (K) and quantification (L–N) of pFoxO1 and FoxO1 (L), MuRF1 (M), and MAFbx (N) in gastrocnemius muscle from male C57BL/6J mice receiving LV-shScrambled, LV-shScrambled + AKG, LV-shADRB2, or LV-shADRB2 + AKG. O, P) Representative images (O) and quantification (P) of propidium iodide (PI, red) positive MuRF1 (green) cells (yellow nucleus indicated by white arrows) in gastrocnemius muscle from male C57BL/6J mice receiving LV-shScrambled, LV-shScrambled + AKG, LV-shADRB2, or LV-shADRB2 + AKG. Q) Levels of 3-MeHis in gastrocnemius muscle from male C57BL/6J mice receiving LV-shScrambled, LV-shScrambled + AKG, LV-shADRB2, or LV-shADRB2 + AKG. Data are presented as means ± sem and were analyzed by nonpaired Student’s t test for panel A, and 1-way ANOVA, followed by post hoc Bonferroni tests for all others. β-Actin served as housekeeping gene. *P < 0.05 compared to control.

Article Snippet: C2C12 cells were ncubated overnight in rabbit anti-PHD3 (bs-0532R; Bioss Antibodies) and rabbit anti-ADRB2 (bs-0947R; Bioss Antibodies) at 4°C.

Techniques: Expressing, Injection, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: 10-Gingerol Enhances the Effect of Taxol in Triple-Negative Breast Cancer via Targeting ADRB2 Signaling

doi: 10.2147/DDDT.S390602

Figure Lengend Snippet: The shRNA Sequence of Lentiviruses

Article Snippet: The following antibodies were used: Bcl-2 (ABCAM, Cambridge, UK, ab182858), Bax (Cell Signaling Technology, Danvers, MA, 2772); cleaved-PARP (Cell Signaling Technology, Danvers, MA, 5625), ADRB2 (Proteintech, Wuhan, China, 13096-1-AP), ERK (ABCAM, Cambridge, UK, ab17942), phospho-ERK1/2 (ABCAM, Cambridge, UK, ab201015), Ki67 (ABCAM, Cambridge, UK, ab16667) and β-actin (Proteintech, Wuhan, China, 66009-1-lg), horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Proteintech, Wuhan, China, SA00001-2), horseradish peroxidase (HRP)-conjugated goat anti-mouse (Proteintech, Wuhan, China, SA00001-1).

Techniques: shRNA, Sequencing, Control

Journal: Drug Design, Development and Therapy

Article Title: 10-Gingerol Enhances the Effect of Taxol in Triple-Negative Breast Cancer via Targeting ADRB2 Signaling

doi: 10.2147/DDDT.S390602

Figure Lengend Snippet: ADRB2 protein is a key target of 10-G action. ( A ) Ligand interaction and binding diagrams of 10-G at ADRB2 active site. ( B ) MDA-MB-231 and SUM-159 cells were transfected with either NC or ADRB2 lentivirus-shRNA for 48 h, and then were treated with different concentrations of 10-G or paclitaxel in combination for 48 h. The cell viability was determined using CCK-8 assay. ( C ) Targeting of ADRB2 with ADRB2 lentivirus-shRNA transfection inhibited the colony formation ability of TNBC cells MDA-MB-231 and SUM-159, and attenuated the cytotoxicity of 10-G induced TNBC cells. Data are represented as the mean value ± SD. *P<0.05, **P<0.01, compared with NC-shRNA.

Article Snippet: The following antibodies were used: Bcl-2 (ABCAM, Cambridge, UK, ab182858), Bax (Cell Signaling Technology, Danvers, MA, 2772); cleaved-PARP (Cell Signaling Technology, Danvers, MA, 5625), ADRB2 (Proteintech, Wuhan, China, 13096-1-AP), ERK (ABCAM, Cambridge, UK, ab17942), phospho-ERK1/2 (ABCAM, Cambridge, UK, ab201015), Ki67 (ABCAM, Cambridge, UK, ab16667) and β-actin (Proteintech, Wuhan, China, 66009-1-lg), horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Proteintech, Wuhan, China, SA00001-2), horseradish peroxidase (HRP)-conjugated goat anti-mouse (Proteintech, Wuhan, China, SA00001-1).

Techniques: Binding Assay, Transfection, shRNA, CCK-8 Assay

Journal: Drug Design, Development and Therapy

Article Title: 10-Gingerol Enhances the Effect of Taxol in Triple-Negative Breast Cancer via Targeting ADRB2 Signaling

doi: 10.2147/DDDT.S390602

Figure Lengend Snippet: 10-G interacts with paclitaxel to inhibit ADRB2/ERK signal. ( A and B ) The protein levels of total ERK and phosphorylated ERK after ADRB2 downregulated by lentivirus-shRNA. ( C and D ) MDA-MB-231 and SUM-159 were treated for 24h with 10-G and paclitaxel alone or in combination, and the protein expression of ADRB2, total ERK and phosphorylated ERK were examined by Western blot. Data are represented as the mean value ± SD. &P<0.05, &&P<0.01, compared with NC-shRNA; **P<0.01, compared with control; +P<0.05, ++P<0.01, compared with 10-G (50μM); ##P<0.01, compared with Paclitaxel; ^P<0.05, ^^P<0.01, compared with Paclitaxel + 10-G (50μM).

Article Snippet: The following antibodies were used: Bcl-2 (ABCAM, Cambridge, UK, ab182858), Bax (Cell Signaling Technology, Danvers, MA, 2772); cleaved-PARP (Cell Signaling Technology, Danvers, MA, 5625), ADRB2 (Proteintech, Wuhan, China, 13096-1-AP), ERK (ABCAM, Cambridge, UK, ab17942), phospho-ERK1/2 (ABCAM, Cambridge, UK, ab201015), Ki67 (ABCAM, Cambridge, UK, ab16667) and β-actin (Proteintech, Wuhan, China, 66009-1-lg), horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Proteintech, Wuhan, China, SA00001-2), horseradish peroxidase (HRP)-conjugated goat anti-mouse (Proteintech, Wuhan, China, SA00001-1).

Techniques: shRNA, Expressing, Western Blot, Control

Journal: Drug Design, Development and Therapy

Article Title: 10-Gingerol Enhances the Effect of Taxol in Triple-Negative Breast Cancer via Targeting ADRB2 Signaling

doi: 10.2147/DDDT.S390602

Figure Lengend Snippet: 10-G enhanced the anti-tumour effect of paclitaxel in vivo. ( A ) Representative tumor image in each treatment group (n=5). ( B ) Tumor volume and tumor weight was measured every 3 days (n=5). ( C ) Representative tumor image of DAPI/TUNEL double-labeling assay of apoptotic cells in each treatment group (n=5). ( D ) The protein levels including Ki67, ADRB2, phosphorylated ERK, Bcl-2, Bax and cleaved PARP was assessed by immunohistochemistry (n=5). Data are represented as the mean value ± SD. **P<0.01, compared with vehicle; ##P<0.01, compared with Paclitaxel. (magnification, ×100).

Article Snippet: The following antibodies were used: Bcl-2 (ABCAM, Cambridge, UK, ab182858), Bax (Cell Signaling Technology, Danvers, MA, 2772); cleaved-PARP (Cell Signaling Technology, Danvers, MA, 5625), ADRB2 (Proteintech, Wuhan, China, 13096-1-AP), ERK (ABCAM, Cambridge, UK, ab17942), phospho-ERK1/2 (ABCAM, Cambridge, UK, ab201015), Ki67 (ABCAM, Cambridge, UK, ab16667) and β-actin (Proteintech, Wuhan, China, 66009-1-lg), horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Proteintech, Wuhan, China, SA00001-2), horseradish peroxidase (HRP)-conjugated goat anti-mouse (Proteintech, Wuhan, China, SA00001-1).

Techniques: In Vivo, TUNEL Assay, Labeling, Immunohistochemistry

Journal: Frontiers in Molecular Biosciences

Article Title: Fingerprinting heterocellular β-adrenoceptor functional expression in the brain using agonist activity profiles

doi: 10.3389/fmolb.2023.1214102

Figure Lengend Snippet: Functional cAMP responses to a panel of β-AR agonists in the C6 rat glioma cell line with (A) native expression, (B) Adrb1 (β 1 -AR) knockout, or (C) Adrb2 (β 2 -AR) knockout. Representative curves are displayed, each with four technical replicates, showing mean ± SEM cAMP HTRF responses normalized to the full agonist isoprenaline. Each agonist curve was converted to a Δlog(E max /EC 50 ) agonist fingerprint using isoprenaline as the reference molecule. Radar plots below the concentration–response curves demonstrate an agonist fingerprint of the C6 cell line non-overlapping with human β 1 -AR or β 2 -AR receptors, suggesting possible species differences and/or dual-receptor expression. (D) Overlay of agonist fingerprints across species for single-receptor systems from either rat (C6 knockout) or human (CHO-K1 recombinant), with an intermediate profile from the C6 co-expressing cell line. (E) Rat primary astrocytes display agonist responses and an agonist fingerprint with hybrid features (F) , indicating endogenous co-expression of β 1 -AR and β 2 -AR.

Article Snippet: ReNcell VM cells expressing β 2 -AR were generated by infecting ReNcell VM cells with a lentivirus containing the ADRB2 gene (OriGene RC204499L3V) at an MOI of 12.5, and 48 h post-infection, cells were grown in media containing 0.25 μg/mL puromycin.

Techniques: Functional Assay, Expressing, Knock-Out, Concentration Assay, Recombinant

Journal: Frontiers in Molecular Biosciences

Article Title: Fingerprinting heterocellular β-adrenoceptor functional expression in the brain using agonist activity profiles

doi: 10.3389/fmolb.2023.1214102

Figure Lengend Snippet: Functional cAMP responses to a panel of β-AR agonists in the human THP-1 cell line with (A) native expression, (B) Adrb1 (β 1 -AR) knockout, or (C) Adrb2 (β 2 -AR) knockout. Representative curves are displayed, each with four technical replicates, showing mean ± SEM cAMP HTRF responses normalized to the full agonist isoprenaline. Each agonist curve was converted to a Δlog(E max /EC 50 ) agonist fingerprint using isoprenaline as the reference molecule. Radar plots below the concentration–response curves reveal an agonist fingerprint of the THP-1 cell line, suggestive of predominantly β 1 -AR function. (D) Overlay of agonist fingerprints across human cell lines with either endogenous or exogenous receptor expression shows good agreement for single-receptor systems. (E) Gene knockout of individual receptors in the THP-1 cell line confirms that the functional cAMP response in parental THP-1 cells is driven predominantly by β 1 -AR receptors; the left subpanel shows cAMP concentration after normalization of the HTRF response.

Article Snippet: ReNcell VM cells expressing β 2 -AR were generated by infecting ReNcell VM cells with a lentivirus containing the ADRB2 gene (OriGene RC204499L3V) at an MOI of 12.5, and 48 h post-infection, cells were grown in media containing 0.25 μg/mL puromycin.

Techniques: Functional Assay, Expressing, Knock-Out, Concentration Assay, Gene Knockout

Journal: Cell

Article Title: Peptidoglycan Production by an Insect-Bacterial Mosaic

doi: 10.1016/j.cell.2019.08.054

Figure Lengend Snippet: A Complete PG Biosynthesis Pathway Is Predicted by Genomics (A) Schematic representation of a single P. citri bacteriocyte (blue), where Moranella cells and their two lipid bilayers (green) reside inside of triple-membrane-bound Tremblaya cells (yellow). (B) Adapted from Typas et al., 2011 . A pictorial representation of the genes present and expressed on the genomes of P. citri (blue represents native eukaryotic genes, brown represents HGTs of Gammaproteobacteria origin, orange represents HGTs of Bacteroidetes origin, and yellow represents Alphaproteobacteria origin) and Moranella (green) that are involved in PG production. The locations of the small colored cell schematics labeled with gene names are based on the predicted location of the protein product of that gene. See also .

Article Snippet: A custom polyclonal antibody was generated (ProSci Incorporated) to a peptide sequence (FVKSLENDYQKTKE) from the P. citri murF gene (accession: AGR65718) that was predicted to be surface exposed.

Techniques: Membrane, Labeling

Journal: Cell

Article Title: Peptidoglycan Production by an Insect-Bacterial Mosaic

doi: 10.1016/j.cell.2019.08.054

Figure Lengend Snippet: Representative Setup for Rearing and Feeding Experiments Involving P. citri , Related to , , , and Mealybugs (yellow arrows) were allowed to feed and reproduce on sprouted potatoes. Where treatments were applied, potatoes (often focusing on the sprouts) were covered and/or injected with 1 mL of the given treatment (modified D- or L-ala, cefsulodin, or water). Mealybugs were carefully placed on the potato with a paintbrush and allowed to eat ad libitum . Treatments were repeated each day for one week.

Article Snippet: A custom polyclonal antibody was generated (ProSci Incorporated) to a peptide sequence (FVKSLENDYQKTKE) from the P. citri murF gene (accession: AGR65718) that was predicted to be surface exposed.

Techniques: Injection, Modification

Journal: Cell

Article Title: Peptidoglycan Production by an Insect-Bacterial Mosaic

doi: 10.1016/j.cell.2019.08.054

Figure Lengend Snippet: MurF, a PG-Related HGT of Alphaproteobacterial Origin, Is Localized to the Moranella Cytoplasm Representative confocal image of a sectioned bacteriome stained with an anti-MurF antibody (red). Insect nuclei are stained with Hoechst (blue). Signal is detected inside of the Moranella cells and insect tissue, but not Tremblaya. Scale bar, 10 μm.

Article Snippet: A custom polyclonal antibody was generated (ProSci Incorporated) to a peptide sequence (FVKSLENDYQKTKE) from the P. citri murF gene (accession: AGR65718) that was predicted to be surface exposed.

Techniques: Staining

Journal: Cell

Article Title: Peptidoglycan Production by an Insect-Bacterial Mosaic

doi: 10.1016/j.cell.2019.08.054

Figure Lengend Snippet:

Article Snippet: A custom polyclonal antibody was generated (ProSci Incorporated) to a peptide sequence (FVKSLENDYQKTKE) from the P. citri murF gene (accession: AGR65718) that was predicted to be surface exposed.

Techniques: Plasmid Preparation, Recombinant, Electron Microscopy, DNA Library Preparation, Amplification, Software