recombinant asprosin  (Elabscience Biotechnology)

Journal: STAR Protocols

Article Title: Overexpression and ELISA-based detection of asprosin in cultured cells and mice

doi: 10.1016/j.xpro.2022.101762

Figure Lengend Snippet: In vitro overexpression of asprosin Asprosin levels measured using ELISA in HEK293T cell conditioned media collected 72 h post transfection with 10 μg of pcDNA3.1 (empty vector control) or pcDNA3.1-IL2sp-6his-Asprosin. Asterisk (∗) indicate the range of alpha; ∗ p<0.05, ∗∗ p<0.01, ∗∗∗ p<0.001, and ∗∗∗∗ p<0.0001, as determined by student T-test. Data presented as mean ± SEM.

Article Snippet: Recombinant human Asprosin , BioLegend , 761902.

Techniques: In Vitro, Over Expression, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation

Journal: STAR Protocols

Article Title: Overexpression and ELISA-based detection of asprosin in cultured cells and mice

doi: 10.1016/j.xpro.2022.101762

Figure Lengend Snippet: Viral overexpression of human asprosin results in increased appetite and body weight gain in lean mice (A and B) cumulative food intake and body weight change were measured 15- and 18-days after 12-week-old, male, C57Bl/6 mice were tail-vein-injected with Ad5-empty or Ad5-h FBN1 (3.6 × 10 9 pfu/mouse) viral vector. (C and D) Cumulative food intake and body weight change were measured 47 days and 60 days after 12-week-old, male, C57Bl/6 mice were tail-vein-injected with AAV8-empty or AAV8-IL2sp-6His-Asprosin (1 × 10 12 GC/mouse) viral vector. Asterisk (∗) indicate the range of alpha; ∗ p<0.05, ∗∗ p<0.01, ∗∗∗ p<0.001, and ∗∗∗∗ p<0.0001, as determined by student T-test. Data presented as mean ± SEM.

Article Snippet: Recombinant human Asprosin , BioLegend , 761902.

Techniques: Over Expression, Injection, Plasmid Preparation

Journal: STAR Protocols

Article Title: Overexpression and ELISA-based detection of asprosin in cultured cells and mice

doi: 10.1016/j.xpro.2022.101762

Figure Lengend Snippet: In vivo overexpression of Asprosin using adenoviral vector (Ad5) (A and B) Human asprosin levels detected in plasma of Ad5-empty (control) and Ad5-h FBN1 injected C57BL/6J male mice, 12 days (A) and 25 days (B) after viral vector transduction. Asprosin detection signal is plotted relative to the average background signal detected in Ad5-empty injected mice. Asterisk (∗) indicate the range of alpha; ∗ p<0.05, ∗∗ p<0.01, ∗∗∗ p<0.001, and ∗∗∗∗ p<0.0001, and (ns) denotes the statistical non significance as determined by student T-test. Data presented as mean ± SEM.

Article Snippet: Recombinant human Asprosin , BioLegend , 761902.

Techniques: In Vivo, Over Expression, Plasmid Preparation, Injection, Transduction

Journal: STAR Protocols

Article Title: Overexpression and ELISA-based detection of asprosin in cultured cells and mice

doi: 10.1016/j.xpro.2022.101762

Figure Lengend Snippet: In vivo overexpression of Asprosin using adeno-associated viral vector 8 (A and B) Human asprosin levels detected in plasma of AAV8-empty (control) and AAV8-IL2sp-6His-Asprosin injected C57BL/6J male mice, 25 days (A) and 55 days (B) after adeno-associated viral vector transduction. Asprosin detection signal is plotted relative to the average background signal detected in AAV8-empty injected mice. Asterisk (∗) indicate the range of alpha; ∗ p<0.05, ∗∗ p<0.01, ∗∗∗ p<0.001, and ∗∗∗∗ p<0.0001, and (ns) denotes the statistical non significance as determined by student T-test. Data presented as mean ± SEM.

Article Snippet: Recombinant human Asprosin , BioLegend , 761902.

Techniques: In Vivo, Over Expression, Plasmid Preparation, Injection, Transduction

Journal: STAR Protocols

Article Title: Overexpression and ELISA-based detection of asprosin in cultured cells and mice

doi: 10.1016/j.xpro.2022.101762

Figure Lengend Snippet:

Article Snippet: Recombinant human Asprosin , BioLegend , 761902.

Techniques: Concentration Assay, Recombinant, Protease Inhibitor, Transfection, Blocking Assay, Cell Culture, Plasmid Preparation

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Asprosin expressions. A, asprosin mRNA and protein in aorta and MA. B, immunofluorescence staining in aorta and MA. Red, asprosin; Green, α-SMA; Blue, DAPI. C, asprosin mRNA and protein in VSMCs. Two-way ANOVA followed by Bonferroni test for A; Unpaired student's test for C. n = 3 or 4. *P < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Immunofluorescence, Staining

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Effects of asprosin overexpression (OE) or knockdown (KD) on related protein expressions in VSMCs. A, asprosin protein. B, NLRP3, ASC, caspase-1, IL-1β protein expressions. Cells were incubated for 48 h with asprosin OE plasmid (2 μg) or asprosin siRNA (50 nM). Two-way ANOVA followed by Bonferroni test. n = 4. *P < 0.05; #P < 0.05 vs WKY.

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Over Expression, Knockdown, Incubation, Plasmid Preparation

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Effects of asprosin protein on NLRP3 inflammasome activation in VSMCs. A, dose-effects of asprosin protein (0. 12.5, 25, 50 and 100 nM) on NLRP3-related protein expressions in WKY and SHR. B, immunofluorescence showing the roles of asprosin (50 nM) on NLRP3 (red) and ASC (green) expressions in SHR. Two-way ANOVA followed by Bonferroni test. n = 4. *P < 0.05 vs 0 nM; #P < 0.05 vs WKY. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Activation Assay, Immunofluorescence

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Asprosin receptors in VSMCs. A, mRNA levels of OR4M1, OLFR734 and TLR4. B and C, effects of asprosin receptor TLR4 KD on asprosin OE-induced NLRP3-related protein expressions. Cells were pretreated with TLR4 siRNA (50 nM) for 24 h before asprosin OE plasmid (2 μg) treatment for 24 h. Two-way ANOVA followed by Bonferroni test. n = 3 or 4. *P < 0.05; #P < 0.05 vs WKY.

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Plasmid Preparation

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Role of NFκB in asprosin overexpression-induced NLRP3 inflammasome activation in VSMCs. A, immunofluorescence staining showing p65-NFκB nuclear translocation. Red, p65-NFκB; Blue, DAPI. B, intracellular and intranuclear p65-NFκB levels. C. Effects of a NFκB inhibitor BAY11-7082 (10 μM) on NLRP3 and IL-1β protein expressions. Two-way ANOVA followed by Bonferroni test. n = 4 or 6. *P < 0.05; #P < 0.05 vs WKY. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Over Expression, Activation Assay, Immunofluorescence, Staining, Translocation Assay

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Effects of NLRP3 inflammasome inhibitor MCC950 (50 μM for 24 h) on asprosin OE-caused proliferation and migration of VSMCs. A-B, VSMC proliferation evaluated with EdU-positive cells (red). C-D, VSMC migration evaluated by Boyden chamber assay. Two-way ANOVA followed by Bonferroni test. n = 6. *P < 0.05; #P < 0.05 vs WKY. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Migration, Boyden Chamber Assay

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Effects of asprosin overexpression in VSMCs from WT and NLRP3 −/− mice. A, NLRP3 mRNA levels. B, NLRP3 and IL-1β protein expressions. C, asprosin protein expression. D-E, VSMC proliferation evaluated with the percentage of EdU-positive cells (red). F-G, VSMC migration evaluated by Boyden chamber assay. Values are mean ± SE. Unpaired student's test for A; Two-way ANOVA followed by Bonferroni test for B–F. n = 6 (A, D, E); n = 4 (B, C). *P < 0.05; #P < 0.05 vs WT. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Over Expression, Expressing, Migration, Boyden Chamber Assay

Journal: Heliyon

Article Title: Asprosin promotes vascular inflammation via TLR4-NFκB-mediated NLRP3 inflammasome activation in hypertension

doi: 10.1016/j.heliyon.2024.e31659

Figure Lengend Snippet: Local asprosin knockdown in common carotid artery attenuates vascular inflammation and remodeling in SHR. A, asprosin expressions in the carotid artery. B, NLRP3 and IL-1β expressions in carotid artery. C, bar graph showing the media thickness, lumen diameter and their ratio in carotid artery. D, hematoxylin-eosin (HE) staining of carotid artery. E, Masson's staining of carotid artery. Two-way ANOVA followed by Bonferroni test. n = 6. *P < 0.05; #P < 0.05 vs WKY.

Article Snippet: Recombinant adenoviruses for asprosin KD and negative control adenoviruses were obtained from Genechem (Shanghai, China).

Techniques: Knockdown, Staining