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: Journal of Cell Communication and Signaling

Article Title: Fibrillin-1 and fibrillin-1-derived asprosin in adipose tissue function and metabolic disorders

doi: 10.1007/s12079-020-00566-3

Figure Lengend Snippet: Furin processing of profibrillin-1 leads to microfibril assembly and asprosin release. Profibrillin-1 is processed by furin cleavage in the N- and C- terminals domains (arrowheads), giving rise to mature fibrillin-1 and the N- and C-terminal propeptides. The C-terminal propeptide (asprosin) is released into the circulation, whereas it is not known whether the N-terminal propeptide fulfils further functions. Processed mature fibrillin-1 assembles to form bead-on-the-string microfibrils. Regions in the fibrillin-1 protein where mutations lead to MFS and MPLS/NPS are indicated by black horizontal bars

Article Snippet: Recently, a group of researchers from Novo Nordisk published a case report describing extensive efforts to reproduce some of the functional aspects of asprosin with many recombinant asprosin constructs produced by bacteria as well as by mammalian cells (Li et al. 2019 ; von Herrath et al. 2019 ).

Techniques:

Journal: Journal of Cell Communication and Signaling

Article Title: Fibrillin-1 and fibrillin-1-derived asprosin in adipose tissue function and metabolic disorders

doi: 10.1007/s12079-020-00566-3

Figure Lengend Snippet: The role of asprosin in metabolic organs. Left solid box: During fasting or in obesity, increased circulating asprosin promotes hepatic glucose production and increases insulin secretion from the pancreas (Romere et al. 2016). Asprosin also activates the AgRP neurons in the brain and increases appetite (Duerrschmid et al. 2017). Both asprosin functions have been tested and validated in vivo and in vitro experiments. Right dashed box: Other in vitro studies have shown that increased asprosin levels lead to apoptosis and inflammation in pancreatic B cells (Lee et al. 2019), and to inflammation and ER stress in the skeletal muscle cells (Jung et al. 2019). However, further in vivo validation is required for these functional aspects

Article Snippet: Recently, a group of researchers from Novo Nordisk published a case report describing extensive efforts to reproduce some of the functional aspects of asprosin with many recombinant asprosin constructs produced by bacteria as well as by mammalian cells (Li et al. 2019 ; von Herrath et al. 2019 ).

Techniques: In Vivo, In Vitro, Biomarker Discovery, Functional Assay

Journal: Journal of Cell Communication and Signaling

Article Title: Fibrillin-1 and fibrillin-1-derived asprosin in adipose tissue function and metabolic disorders

doi: 10.1007/s12079-020-00566-3

Figure Lengend Snippet: Summary of the principle roles of asprosin in different metabolic tissues and organs

Article Snippet: Recently, a group of researchers from Novo Nordisk published a case report describing extensive efforts to reproduce some of the functional aspects of asprosin with many recombinant asprosin constructs produced by bacteria as well as by mammalian cells (Li et al. 2019 ; von Herrath et al. 2019 ).

Techniques: Binding Assay

Journal: Nature medicine

Article Title: Asprosin is a centrally-acting orexigenic hormone

doi: 10.1038/nm.4432

Figure Lengend Snippet: Introducing the NPS mutation in mice results in hypophagia, reduced adiposity and protection from diet-induced obesity. ( a ) Schematic depiction of the CRISPR/Cas9 strategy employed to generate Fbn1 NPS/+ mice. A small (10 bp) deletion was introduced at the junction of exon 65 and intron 65 (top left), resulting in loss of a splice site, leading to skipping of exon 65 (middle left) and truncation of profibrillin (bottom left), identical to the molecular events in an individual with NPS (WT and heterozygous mutant sequence trace, WT and mutant mRNA sequence at the deletion site, and resulting WT and mutant protein sequence – top, middle, and bottom right, respectively). ( b ) Sandwich Elisa for endogenous asprosin in plasma of WT and Fbn1 NPS/+ mice (WT n = 6, NPS n = 7). ( c ) Photograph of a representative set ( n = 12 for each group) of 5-month-old male WT mice and Fbn1 NPS/+ littermates on a high-fat diet for 3 months. ( d , e ) Body composition data using DEXA scans for d WT and Fbn1 NPS/+ mice on normal chow (WT n = 8, NPS n = 7) and e for WT and Fbn1 NPS/+ mice on a high-fat diet for 3 months ( n = 8 per group). ( f ) Weight curves of WT and Fbn1 NPS/+ mice 4 to 14 weeks old ( n = 6 per group, P = 0.008). ( g ) Cumulative food intake over 24 hr in mice from d in the ad libitum fed and overnight fasted state using the CLAMs system. ( h ) Energy expenditure was measured over 24 hr mice from d using the CLAMs system ( P = 0.16). ( i ) Analysis of energy expenditure of Fbn1 NPS/+ mice and WT littermates from d on normal chow by ANCOVA ( n = 5 per group, body weight P = 0.009, lean mass P = 0.016). ( j ) Firing frequency and membrane potential of AgRP + neurons from ad libitum fed and over-night fasted WT and Fbn1 NPS/+ mice (Firing frequency: fed, WT n = 24; fed, NPS n = 23; fasted, WT n = 25; fasted, NPS n = 20. Membrane potential: fed, WT n = 24; fed, NPS n = 23; fasted, WT n = 25; fasted, NPS n = 28). * P < 0.05, ** P < 0.01, and *** P < 0.001. Statistical tests used: two-tailed t -test ( b,d,e ) or two-way ANOVA ( f,g,h,j ).

Article Snippet: To obtain cerebrospinal fluid (CSF) for asprosin analysis, we contracted BioreclamationIVT to inject 8- to 12-week old male rats intravenously with His-tagged recombinant asprosin followed by sacrificing them and obtaining CSF 1 hr after injection.

Techniques: Mutagenesis, CRISPR, Sequencing, Sandwich ELISA, Clinical Proteomics, Membrane, Two Tailed Test

Journal: Nature medicine

Article Title: Asprosin is a centrally-acting orexigenic hormone

doi: 10.1038/nm.4432

Figure Lengend Snippet: Correcting the asprosin deficiency completely rescues hypophagia and depressed AgRP + neuron activity in Fbn1 NPS/+ mice. ( a ) Cumulative food intake over 24 hr in WT and Fbn1 NPS/+ mice after subcutaneous injection of recombinant GFP or mammalian-expressed recombinant asprosin using the CLAMs system (30 μg/mouse, n = 5 per group). ( b ) Membrane potential and firing frequency of AgRP + neurons from overnight fasted WT and Fbn1 NPS/+ mice 3 hr after ICV injection of recombinant GFP or mammalian-expressed recombinant asprosin as indicated (rAsprosin: 0.5 μg; rGFP: 0.5 μg; membrane potential WT + rGFP: n = 35, NPS + rGFP: n = 31, NPS + rAsprosin: n = 20; firing frequency: WT + rGFP: n = 25, NPS + rGFP: n = 28, NPS + rAsprosin: n = 18). ( c ) Membrane potential and firing frequency of AgRP + neurons from overnight fasted WT and Fbn1 NPS/+ mice after 2 hr of incubation of intact hypothalamic slices with recombinant GFP or bacterially expressed recombinant asprosin as indicated (rAsprosin: 34 nM; rGFP: 0.5 μg/μL; membrane potential: WT + rGFP: n = 12, NPS + rGFP: n = 14, NPS + rAsprosin: n = 14; firing frequency: WT + rGFP: n = 12, NPS + rGFP: n = 11, NPS + rAsprosin: n = 13). * P < 0.05, and *** P < 0.001. Statistical tests used: one-way ANOVA ( a–c ).

Article Snippet: To obtain cerebrospinal fluid (CSF) for asprosin analysis, we contracted BioreclamationIVT to inject 8- to 12-week old male rats intravenously with His-tagged recombinant asprosin followed by sacrificing them and obtaining CSF 1 hr after injection.

Techniques: Activity Assay, Injection, Recombinant, Membrane, Incubation

Journal: Nature medicine

Article Title: Asprosin is a centrally-acting orexigenic hormone

doi: 10.1038/nm.4432

Figure Lengend Snippet: Asprosin crosses the blood-brain-barrier and stimulates appetite ( a ) Endogenous asprosin in cerebrospinal fluid of ad libitum fed and over-night fasted rats using a sandwich Elisa ( n = 7 per group). ( b ) N-terminal His-tag (on bacterially expressed asprosin) and total asprosin (recombinant + endogenous) in cerebrospinal fluid of fasted rats after intravenous injection of bacterially expressed, His-tagged asprosin using a sandwich Elisa (30 μg/mouse, n = 4 per group). ( c ) Food intake during 24 hr after a single subcutaneous injection of recombinant GFP or bacterially expressed asprosin in mice using the CLAMs system (30 μg/mouse, n = 5 per group, P = 0.06). ( d ) Food intake during 24 hr after a single subcutaneous injection of recombinant GFP or mammalian-expressed recombinant asprosin in mice using the CLAMs system (60 μg/mouse, n = 6 per group, P = 0.0003). ( e ) Cumulative food intake during the dark phase (12 hr) of circadian entrained mice after intracerebroventricular (ICV) injection of recombinant GFP or bacterially expressed recombinant asprosin (rAsprosin: 10 ng; rGFP: 10 ng, n = 8 per group). ( f ) Cumulative food intake over 24 hr in mice exposed to 10 days of a single daily injection of recombinant GFP or bacterially expressed asprosin using the CLAMs system (30 μg/mouse/day, n = 5 per group). ( g ) Energy expenditure over 24 hr in mice from f using the CLAMs system ( P = 0.15). ( h ) Fat mass from magnetic resonance imaging (MRI) in mice before and after 10 days of a single daily injection of recombinant GFP or bacterially expressed recombinant asprosin in mice from f . ( i ) Cumulative food intake over 24 hr in mice 10 days after adenoviral overexpression of GFP or FBN1 using the CLAMs system ( n = 5 per group). ( j ) Energy expenditure over 24 hr in mice from (i) using the CLAMs system ( P = 0.46). ( k ) Fat mass from MRI in mice from i before and 10 days after injection with the GFP or FBN1 adenovirus ( n = 5 per group, P = 0.06 between FBN1 group 0 and 10 days). ( l ) Fat mass from MRI in mice 1 week and 3 weeks after injection with the GFP or FBN1 adenovirus ( n = 5 per group). * P < 0.05, and ** P < 0.01. Statistical tests used: two-tailed t -test ( a,b,e,f,i ) or two-way ANOVA ( c,d,g,h,j,k,l ).

Article Snippet: To obtain cerebrospinal fluid (CSF) for asprosin analysis, we contracted BioreclamationIVT to inject 8- to 12-week old male rats intravenously with His-tagged recombinant asprosin followed by sacrificing them and obtaining CSF 1 hr after injection.

Techniques: Sandwich ELISA, Recombinant, Injection, Magnetic Resonance Imaging, Over Expression, Two Tailed Test

Journal: Nature medicine

Article Title: Asprosin is a centrally-acting orexigenic hormone

doi: 10.1038/nm.4432

Figure Lengend Snippet: AgRP + neurons are essential for Asprosin-mediated appetite stimulation ( a ) Representative action potential (AP) firing traces of AgRP + neurons after GFP and bacterially expressed recombinant asprosin treatment. ( b ) Response ratio of AgRP + neurons after GFP, and 1 nM and 34 nM bacterially expressed asprosin treatment (rGFP n = 8, 1 nM rAsprosin n = 12, 34 nM rAsprosin n = 46). ( c ) Representative traces of miniature excitatory postsynaptic current (mEPSC) in AgRP + neurons before and after bacterially expressed asprosin treatment. ( d ) mEPSC frequency and amplitude in AgRP + neurons before and after bacterially expressed asprosin treatment ( n = 6 per group). ( e ) Representative traces of AgRP + neuron resting membrane potential in the presence of TTX (1 μM; top), and inhibitor cocktail (AP-5: 30 μM, CNQX:30 μM, bicuculline: 50 μM and TTX 1 μM; bottom). ( f ) Amplitude changes of resting membrane potential in AgRP + neurons after treatment with GFP, 1 nM and 34 nM bacterially expressed asprosin, or 34 nM bacterially expressed asprosin in the presence of TTX and inhibitor cocktail (AP-5: 30 μM, CNQX: 30 μM, bicuculline: 50 μM, and TTX 1 μM; rGFP n = 8, rAsprosin 1 nM n = 12, 34 nM n = 44, 34 nM + TTX n = 11, 34 nM + TTX + CNQX + AP5 + bicuculline n = 13). ( g ) Response ratio of AgRP + neurons after treatment with bacterially expressed asprosin in the presence of TTX or inhibitor cocktail (AP-5: 30 μM, CNQX: 30 μM, bicuculline: 50 μM, and TTX 1 μM; rAsprosin + TTX n = 11, rAsprosin + inhibitors n = 13). ( h ) Cumulative food intake on normal-chow in WT and AgRP + neuron-ablated mice in response to a single dose of GFP or bacterially expressed asprosin (30 μg/mouse, n = 4 per group). * P < 0.05. Statistical tests used: two-tailed t -test ( e ), one-way ANOVA ( f ), or two-way ANOVA ( h ).

Article Snippet: To obtain cerebrospinal fluid (CSF) for asprosin analysis, we contracted BioreclamationIVT to inject 8- to 12-week old male rats intravenously with His-tagged recombinant asprosin followed by sacrificing them and obtaining CSF 1 hr after injection.

Techniques: Recombinant, Membrane, Two Tailed Test

Journal: Nature medicine

Article Title: Asprosin is a centrally-acting orexigenic hormone

doi: 10.1038/nm.4432

Figure Lengend Snippet: Immunologic neutralization of asprosin is protective against obesity ( a ) Firing frequency and membrane potential in AgRP + neurons from mice 12 hr after IgG control or anti-asprosin monoclonal antibody injection (250 μg/mouse, firing frequency: IgG n = 27, anti-asprosin mAb n = 26; membrane potential: IgG n = 33, anti-asprosin mAb n = 34). ( b ) Cumulative food intake over 24 hr in 8-week-old WT mice with one daily injection of IgG control or anti-asprosin mAb in the ad libitum fed and overnight fasted state using the CLAMs system (250 μg/mouse, n = 5 per group). ( c ) Energy expenditure over 24 hr in ad libitum fed mice from b using the CLAMs system ( P = 0.4). ( d ) Cumulative food intake over 24 hr in 8-week-old male Lepr db/db mice with one daily injection IgG control or anti-asprosin mAb using the CLAMs system (250 μg/mouse, n = 5 per group). ( e ) Energy expenditure over 24 hr in mice from (d) using the CLAMs system ( P = 0.22). ( f ) Weight change over time in mice from d and littermates ( n = 6 per group, P = 0.007). Original weights: 43.8 g for IgG group and 46.9 g for anti-asprosin mAb group. ( g ) Cumulative food intake during 24 hr in 20-week-old male WT mice fed a high-fat diet for 3 months with daily dosing for 5 days of IgG control or anti-asprosin mAb using the CLAMS system (250 μg/mouse, n = 5 per group). ( h ) Energy expenditure over 24 hr in mice from g using the CLAMs system ( P = 0.67). ( i ) Weight change over time in mice from g and littermates with daily dosing for 5 days with IgG control or anti-asprosin mAb (250 μg/mouse, n = 6 per group, P = 0.05). Original weights: 50.1 g for IgG group and 50.3 g for anti-asprosin mAb group. * P < 0.05, and ** P < 0.01. Statistical tests used: two-tailed t -test ( a,d,g ) or two-way ANOVA ( b,c,e,f,h,i ).

Article Snippet: To obtain cerebrospinal fluid (CSF) for asprosin analysis, we contracted BioreclamationIVT to inject 8- to 12-week old male rats intravenously with His-tagged recombinant asprosin followed by sacrificing them and obtaining CSF 1 hr after injection.

Techniques: Neutralization, Membrane, Control, Injection, Two Tailed Test

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

Journal: eLife

Article Title: Asprosin-neutralizing antibodies as a treatment for metabolic syndrome

doi: 10.7554/eLife.63784

Figure Lengend Snippet: Markers of general sickness (half hourly pedestrian activity and wheel running activity), plasma levels of pro-inflammatory (IL1β and TNFα), and anti-inflammatory (IL10 and TGFβ) cytokines, markers of renal (plasma creatinine and blood urea nitrogen [BUN] levels) and hepatic health (plasma ALT, alanine aminotransferase and AST, aspartate aminotransferase levels) were measured in 16-week-old diet-induced obese (DIO) mice intra-peritoneally injected with control, isotype-matched IgG, or anti-asprosin mAb (250 µg/mouse). Downward arrow indicates the time of antibody treatment in the top panel.

Article Snippet: Pro-inflammatory (IL1β [Abcam, catalog # ab197742] and TNFα [Abcam, catalog # ab208348]) and anti-inflammatory (IL10 [Thermo Scientific, catalog # BMS614INST] and TGFβ [R and D systems, catalog #DB100B]) cytokines were measured in plasma collected from 16-week-old male DIO mice intra-peritoneally injection of control, isotype-matched IgG, or anti-asprosin mouse mAb (250 µg mAb in 500 µl 0.9% saline).

Techniques: Activity Assay, Injection

Journal: eLife

Article Title: Asprosin-neutralizing antibodies as a treatment for metabolic syndrome

doi: 10.7554/eLife.63784

Figure Lengend Snippet:

Article Snippet: Pro-inflammatory (IL1β [Abcam, catalog # ab197742] and TNFα [Abcam, catalog # ab208348]) and anti-inflammatory (IL10 [Thermo Scientific, catalog # BMS614INST] and TGFβ [R and D systems, catalog #DB100B]) cytokines were measured in plasma collected from 16-week-old male DIO mice intra-peritoneally injection of control, isotype-matched IgG, or anti-asprosin mouse mAb (250 µg mAb in 500 µl 0.9% saline).

Techniques: Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Quantitation Assay