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FIGURE 6 Shensu IV regulates the PI3K/AKT signaling pathway through H2S. (A) The effects of Shensu IV and NaHS on the mRNA expression <t>of</t> <t>CD2AP,</t> nephrin, <t>CBS,</t> CSE, NOX4, PI3K, and AKT in renal tissue of PAN rats were analyzed by RT-qPCR. (B) Western blot analysis of the effects of Shensu IV and NaHS on the protein levels of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, p-PI3K,AKT,p-AKT in renal tissue of PAN rats. *P< 0.05, **P< 0.01, ***P< 0.001. Abbreviations: CD2AP, CD2-associated protein; CBS, Cystathionine β-synthase; CSE, Cystathionine γ-lyase; PI3K, Phosphoinositide 3-Kinase; AKT, Protein Kinase B.

Cbs, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 13 article reviews

cbs - by Bioz Stars, 2026-09

92/100 stars

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Article Title: Shensu IV maintains the integrity of the glomerular filtration barrier and exerts renal protective effects by regulating endogenous hydrogen sulfide levels

Journal: Frontiers in Pharmacology

doi: 10.3389/fphar.2024.1447249

FIGURE 6 Shensu IV regulates the PI3K/AKT signaling pathway through H2S. (A) The effects of Shensu IV and NaHS on the mRNA expression of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, and AKT in renal tissue of PAN rats were analyzed by RT-qPCR. (B) Western blot analysis of the effects of Shensu IV and NaHS on the protein levels of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, p-PI3K,AKT,p-AKT in renal tissue of PAN rats. *P< 0.05, **P< 0.01, ***P< 0.001. Abbreviations: CD2AP, CD2-associated protein; CBS, Cystathionine β-synthase; CSE, Cystathionine γ-lyase; PI3K, Phosphoinositide 3-Kinase; AKT, Protein Kinase B.

Figure Legend Snippet: FIGURE 6 Shensu IV regulates the PI3K/AKT signaling pathway through H2S. (A) The effects of Shensu IV and NaHS on the mRNA expression of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, and AKT in renal tissue of PAN rats were analyzed by RT-qPCR. (B) Western blot analysis of the effects of Shensu IV and NaHS on the protein levels of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, p-PI3K,AKT,p-AKT in renal tissue of PAN rats. *P< 0.05, **P< 0.01, ***P< 0.001. Abbreviations: CD2AP, CD2-associated protein; CBS, Cystathionine β-synthase; CSE, Cystathionine γ-lyase; PI3K, Phosphoinositide 3-Kinase; AKT, Protein Kinase B.

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

FIGURE 8 Shensu IV regulates the PI3K/AKT signaling pathway through H2S in podocytes. (A) The effects of Shensu IV and NaHS on the mRNA expression of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, and AKT in podocytes were analyzed by RT-qPCR. (B) Western blot analysis of the effects of Shensu IV and NaHS on the protein levels of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, p-PI3K,AKT,p-AKT in PAN-induced podocyocytes. *P< 0.05, **P< 0.01, ***P< 0.001. Abbreviations: CD2AP, CD2-associated protein; CBS, Cystathionine β-synthase; CSE, Cystathionine γ-lyase; PI3K, Phosphoinositide 3-Kinase; AKT, Protein Kinase B.

Figure Legend Snippet: FIGURE 8 Shensu IV regulates the PI3K/AKT signaling pathway through H2S in podocytes. (A) The effects of Shensu IV and NaHS on the mRNA expression of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, and AKT in podocytes were analyzed by RT-qPCR. (B) Western blot analysis of the effects of Shensu IV and NaHS on the protein levels of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, p-PI3K,AKT,p-AKT in PAN-induced podocyocytes. *P< 0.05, **P< 0.01, ***P< 0.001. Abbreviations: CD2AP, CD2-associated protein; CBS, Cystathionine β-synthase; CSE, Cystathionine γ-lyase; PI3K, Phosphoinositide 3-Kinase; AKT, Protein Kinase B.

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

Blocking Assay:

Article Title: Rbm20 ablation is associated with changes in the expression of titin-interacting and metabolic proteins.
Article Snippet: Proteins were transferred to Immun-Blot PVDF Membranes for Protein Blotting (0.2 mm pore size, Bio-Rad, cat# 1620177) at 300 mA for 90 min in a cold room (4 1C). .. To block, membranes were incubated in TBST with 5% (w/v) nonfat dry milk for 1 h at room temperature, followed by incubation in primary antibody solution containing 5% (w/v) nonfat dry milk and diluted Msrb2 (1 : 500, Proteintech, cat# 17629-1-AP) or Gapdh (1 : 1000, Cell Signaling Technology, cat# 2118) primary antibodies in TBST overnight at 4 1C. .. The membranes were removed from primary antibody solution and washed 5 5 min with TBST followed by incubation in TBST with 3% (w/v) nonfat dry milk and diluted HRP-conjugated secondary antibody (1 : 3000, Promega, cat# W4018) for 1 h at room temperature.

Article Title: Effect of Cr content on microstructure and mechanical properties of annealed large-dimensional bulk nanocrystalline Fe–Al–Cr alloys by self-propagating combustion synthesis
Article Snippet: BACKGROUND: Methionine is an essential sulfur-containing amino acid.. To elucidate the influence of L-methionine on activation of the nuclear factor erythroid 2-related factor 2–antioxidant responsive element (Nrf2-ARE) antioxidant pathway to stimulate the endogenous antioxidant activity for depressing reactive oxygen species (ROS)-derived oxidative stress, male Wistar rats were orally administered L-methionine daily for 14 days.. RESULTS: With the intake of L-methionine, Nrf2 was activated by L-methionine through depressing Keap1 and Cul3, resulting in upregulation of ARE-driven antioxidant expression (glutamate cysteine ligase catalytic subunit, glutamate cysteine ligase modulatory subunit, glutathione synthase (GS), catalase (CAT), superoxide dismutase (SOD), heme oxygenase 1, NAD(P)H:quinone oxidoreductase 1, glutathione reductase (GR), glutathione S-transferase (GST), glutathione peroxidase (GPx)) with increasing L-methionine availability.

Article Title: Methionine augments endogenous antioxidant capacity of rice protein through stimulating MSR antioxidant system and activating Nrf2-ARE pathway in growing and adult rats
Article Snippet: To elucidate the influence of methionine on the endogenous antioxidant activity of rice protein (RP), growing and adult rats were fed with RP and methionine-supplemented RP (RM).. After 2 weeks feeding, hepatic contents of ROS were significantly reduced by RP and RM.. The endogenous antioxidant responses were induced by RP and increased by RM, in which methionine sulfoxide reductases (MsrA, MsrB2, MsrB3) expression and glutathione synthesis were uniformly stimulated and up-regulated with increasing consumption of methionine.

Incubation:

Protein Extraction:

Article Title: Rice Protein Exerts Endogenous Antioxidant Capacity via Methionine Sulfoxide Reductase and the Nrf2 Antioxidant System Independent of Age.
Article Snippet: .. Protein extraction and Western blotting analysis The total, cytoplasmic, and nuclear proteins were used for Western blot analysis, which were prepared as described in our previous studies.16,22,23,28 The primary antibodies of Nrf2, Kelch-like ECH-associated protein 1 (Keap1), Cullin 3 (Cul3), MsrA, MsrB2, MsrB3, glutamate cysteine ligase catalytic subunit (GCLC), glutamate cysteine ligase modulatory subunit (GCLM), glutathione synthase (GS), GR, GSTA1, SOD, CAT, heme oxygenase 1 (HO-1), NAD(P)H: quinone oxidoreductase 1 (NQO1), H1.2, and GAPDH were purchased from Proteintech (Wuhan, China), and methionine adenosyltransferase Ia (MAT1A), S-adenosylhomocysteine hydrolase (SAHH), cystathionine b-synthase (CBS), cystathionine c-lyse (CTH), GPx1/2, and second antibody were Table 1. ..

Western Blot:

Saline:

Image Search Results

a , (Top) Homochirality paradigm present within the biomacromolecules of life. (Middle) Chiral post-translational modification of Met and native Met redox regulation. (Bottom) ChURRO strategy for the global detection of prochiral Met oxidation sites. b , Development of structurally diverse ChURRO library identifies robust ChURRO probes with high methionine labeling in HEK-293T lysates. c , Msr rescue of oxidized proteins and labeling by ChURRO-2 using purified MsrA and MsrB enzymes and target substrates in vitro . d , ChURRO-ABPP strategy to validate detection of prochiral Met oxidation sites. HEK-293T cells were incubated with siRNA (50 nM) to knock-down endogenous MsrB2 or MsrA followed by addition of paraquat (100 µM) to bias ( R )- or ( S )-MetO formation on proteins. Following lysis, lysates were treated with equimolar (200 µM) doses of ( S )- or ( R )-ChURRO-2 and labeled with isotopically-encoded desthiobiotin tags through Cu-catalyzed click chemistry. Proteomes were then combined, enriched using streptavidin, tryptically digested, and analyzed by LC-MS/MS. Volcano plots of ChURRO-ABPP identifying prochiral Met oxidation sites in oxidatively-stressed HEK-293T lysates lacking either ( R )- or ( S )-MetO rescue mechanisms were then generated (n = 12 technical MS replicates).

Journal: bioRxiv

Article Title: Chiral methionine oxidation reagents reveal stereospecific proteome modifications

doi: 10.64898/2026.03.24.713977

Figure Lengend Snippet: a , (Top) Homochirality paradigm present within the biomacromolecules of life. (Middle) Chiral post-translational modification of Met and native Met redox regulation. (Bottom) ChURRO strategy for the global detection of prochiral Met oxidation sites. b , Development of structurally diverse ChURRO library identifies robust ChURRO probes with high methionine labeling in HEK-293T lysates. c , Msr rescue of oxidized proteins and labeling by ChURRO-2 using purified MsrA and MsrB enzymes and target substrates in vitro . d , ChURRO-ABPP strategy to validate detection of prochiral Met oxidation sites. HEK-293T cells were incubated with siRNA (50 nM) to knock-down endogenous MsrB2 or MsrA followed by addition of paraquat (100 µM) to bias ( R )- or ( S )-MetO formation on proteins. Following lysis, lysates were treated with equimolar (200 µM) doses of ( S )- or ( R )-ChURRO-2 and labeled with isotopically-encoded desthiobiotin tags through Cu-catalyzed click chemistry. Proteomes were then combined, enriched using streptavidin, tryptically digested, and analyzed by LC-MS/MS. Volcano plots of ChURRO-ABPP identifying prochiral Met oxidation sites in oxidatively-stressed HEK-293T lysates lacking either ( R )- or ( S )-MetO rescue mechanisms were then generated (n = 12 technical MS replicates).

Article Snippet: Separate producer cells were transfected using 1 μg of lentiviral plasmids psPAX2 and pMD2.G each as well as 3 μg of plasmid containing lentiCRISPR v2-sgRNA plasmid bearing either MsrA g1/g2, or MsrB2 g1/g2 following the TransIT-293 transfection protocol (Mirusbio; MIR 2700).

Techniques: Modification, Labeling, Purification, In Vitro, Incubation, Knockdown, Lysis, Liquid Chromatography with Mass Spectroscopy, Generated

a , Homocysteine thiolactone (HCTL) regulation by BPHL and methionyl–tRNA synthetase (MRS) and subsequent protein N -homocysteinylation with HCTL. b , RoseTTAFold All-Atom structure of human BPHL with modeled ( R )-MetO at the 69 site. c , k M of BPHL variants under conditions of no stress or oxidative stress (n = 3 technical replicates, error bars represent standard deviation). d , Dose-response curve of WT BPHL treated with ( S )- and ( R )-ChURRO-2 with calculated IC 50 values (n = 3 technical replicates, error bars represent standard deviation). e , LC-MS/MS quantification of 18 O-H 2 O 2 adducts at BPHL M69 with MsrB2 treatment (n = 3 technical MS replicates, error bars represent standard deviation).

Journal: bioRxiv

Article Title: Chiral methionine oxidation reagents reveal stereospecific proteome modifications

doi: 10.64898/2026.03.24.713977

Figure Lengend Snippet: a , Homocysteine thiolactone (HCTL) regulation by BPHL and methionyl–tRNA synthetase (MRS) and subsequent protein N -homocysteinylation with HCTL. b , RoseTTAFold All-Atom structure of human BPHL with modeled ( R )-MetO at the 69 site. c , k M of BPHL variants under conditions of no stress or oxidative stress (n = 3 technical replicates, error bars represent standard deviation). d , Dose-response curve of WT BPHL treated with ( S )- and ( R )-ChURRO-2 with calculated IC 50 values (n = 3 technical replicates, error bars represent standard deviation). e , LC-MS/MS quantification of 18 O-H 2 O 2 adducts at BPHL M69 with MsrB2 treatment (n = 3 technical MS replicates, error bars represent standard deviation).

Techniques: Standard Deviation, Liquid Chromatography with Mass Spectroscopy

a , Mitochondrial uncoupler assay to assess protein N -homocysteinylation status as a function of BPHL Met redox status using thioester probe AT-3 to label sites of protein N -homocysteinylation. b , BPHL and Msr co-immunoprecipitation analysis following mitochondrial uncoupling in HEK-293T cells (n = 3 cell technical replicates, error bars represent standard deviation). c , Relative protein N -homocysteinylation levels following loss of mitochondrial ( S )- or ( R )-MetO rescue mechanisms (n = 3 cell technical replicates, error bars represent standard deviation). d , Chemoproteomic scheme for assessing global N -homocysteinylation changes in MsrB2 deficient cell models. HEK-293T cells were incubated with siRNA (50 nM) to knock-down endogenous MsrB2 addition of paraquat (100 μM) and HCTL (500 μM). Following lysis, lysates were treated with equimolar (1 mM) doses of AT-3 and labeled with isotopically-encoded desthiobiotin tags through Cu-catalyzed click chemistry. Proteomes were then combined, enriched using streptavidin, tryptically digested, and analyzed by LC-MS/MS. e , Volcano plot of hyper N -homocysteinylated sites detected in oxidatively-stressed HEK-293T lysates containing or lacking ( R )-MetO rescue mechanisms (n = 4 technical MS replicates). f , (Left) Alphafold2 model of SOD1 with K76 site shown. (Right) Cell activity assays of HEK-293T cells expressing wild-type or K76R SOD1 upon titration of HCTL (n = 3 cell technical replicates, error bars represent standard deviation).

Journal: bioRxiv

Article Title: Chiral methionine oxidation reagents reveal stereospecific proteome modifications

doi: 10.64898/2026.03.24.713977

Figure Lengend Snippet: a , Mitochondrial uncoupler assay to assess protein N -homocysteinylation status as a function of BPHL Met redox status using thioester probe AT-3 to label sites of protein N -homocysteinylation. b , BPHL and Msr co-immunoprecipitation analysis following mitochondrial uncoupling in HEK-293T cells (n = 3 cell technical replicates, error bars represent standard deviation). c , Relative protein N -homocysteinylation levels following loss of mitochondrial ( S )- or ( R )-MetO rescue mechanisms (n = 3 cell technical replicates, error bars represent standard deviation). d , Chemoproteomic scheme for assessing global N -homocysteinylation changes in MsrB2 deficient cell models. HEK-293T cells were incubated with siRNA (50 nM) to knock-down endogenous MsrB2 addition of paraquat (100 μM) and HCTL (500 μM). Following lysis, lysates were treated with equimolar (1 mM) doses of AT-3 and labeled with isotopically-encoded desthiobiotin tags through Cu-catalyzed click chemistry. Proteomes were then combined, enriched using streptavidin, tryptically digested, and analyzed by LC-MS/MS. e , Volcano plot of hyper N -homocysteinylated sites detected in oxidatively-stressed HEK-293T lysates containing or lacking ( R )-MetO rescue mechanisms (n = 4 technical MS replicates). f , (Left) Alphafold2 model of SOD1 with K76 site shown. (Right) Cell activity assays of HEK-293T cells expressing wild-type or K76R SOD1 upon titration of HCTL (n = 3 cell technical replicates, error bars represent standard deviation).

Techniques: Immunoprecipitation, Standard Deviation, Incubation, Knockdown, Lysis, Labeling, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Expressing, Titration

a , BPHL expression screen in renal and hepatic cell lines assessed by immunoblotting (n = 3 technical cell replicates, error bars represent standard deviation). b , Immunoprecipitation studies of FLAG-BPHL expressed in HEK-293T cells using ( R )- and ( S )-ChURRO-2. c , Representative immunoblot of BPHL Co-IP with MsrA and MsrB2 (n = 3 cell technical replicates). d , Immunoblot analysis of MsrA and MsrB2 KO efficiency. e , In-gel fluorescence data of labeled N -homocysteinylated proteins in HEK-293T cells with MsrA or MsrB2 KO (n = 3 technical cell replicates). f , Assessment of Msr knock-down efficiency in HEK-293T cells.

Journal: bioRxiv

Article Title: Chiral methionine oxidation reagents reveal stereospecific proteome modifications

doi: 10.64898/2026.03.24.713977

Figure Lengend Snippet: a , BPHL expression screen in renal and hepatic cell lines assessed by immunoblotting (n = 3 technical cell replicates, error bars represent standard deviation). b , Immunoprecipitation studies of FLAG-BPHL expressed in HEK-293T cells using ( R )- and ( S )-ChURRO-2. c , Representative immunoblot of BPHL Co-IP with MsrA and MsrB2 (n = 3 cell technical replicates). d , Immunoblot analysis of MsrA and MsrB2 KO efficiency. e , In-gel fluorescence data of labeled N -homocysteinylated proteins in HEK-293T cells with MsrA or MsrB2 KO (n = 3 technical cell replicates). f , Assessment of Msr knock-down efficiency in HEK-293T cells.

Techniques: Expressing, Western Blot, Standard Deviation, Immunoprecipitation, Co-Immunoprecipitation Assay, Fluorescence, Labeling, Knockdown

Journal: Frontiers in Pharmacology

Article Title: Shensu IV maintains the integrity of the glomerular filtration barrier and exerts renal protective effects by regulating endogenous hydrogen sulfide levels

doi: 10.3389/fphar.2024.1447249

Figure Lengend Snippet: FIGURE 6 Shensu IV regulates the PI3K/AKT signaling pathway through H2S. (A) The effects of Shensu IV and NaHS on the mRNA expression of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, and AKT in renal tissue of PAN rats were analyzed by RT-qPCR. (B) Western blot analysis of the effects of Shensu IV and NaHS on the protein levels of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, p-PI3K,AKT,p-AKT in renal tissue of PAN rats. *P< 0.05, **P< 0.01, ***P< 0.001. Abbreviations: CD2AP, CD2-associated protein; CBS, Cystathionine β-synthase; CSE, Cystathionine γ-lyase; PI3K, Phosphoinositide 3-Kinase; AKT, Protein Kinase B.

Article Snippet: Membranes were blocked with 5% skim milk (Solarbio) to prevent nonspecific binding and then incubated with primary antibodies against CD2AP (1:2000; A01756-2, BOSTER, Wuhan, China), nephrin (1:2000; A01756-2, BOSTER), CBS (1: 10,000; 14787-1-AP, Proteintech, Wuhan, China), CSE (1: 4,000; 12217-1-AP, Proteintech), PI3K (1:2000; 60225-1-Ig, Proteintech), p-PI3K (1:2000; bs-3332R, BOSTER), AKT (1:10,000; 60203-2-Ig, Proteintech), p-AKT (1: 10,000; 66,444-1-lg, Proteintech), NOX4 (1: 8,000; 14347-1-AP, Proteintech), and GAPDH (1:40,000; 60004-1- Ig, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Shensu IV maintains the integrity of the glomerular filtration barrier and exerts renal protective effects by regulating endogenous hydrogen sulfide levels

doi: 10.3389/fphar.2024.1447249

Figure Lengend Snippet: FIGURE 8 Shensu IV regulates the PI3K/AKT signaling pathway through H2S in podocytes. (A) The effects of Shensu IV and NaHS on the mRNA expression of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, and AKT in podocytes were analyzed by RT-qPCR. (B) Western blot analysis of the effects of Shensu IV and NaHS on the protein levels of CD2AP, nephrin, CBS, CSE, NOX4, PI3K, p-PI3K,AKT,p-AKT in PAN-induced podocyocytes. *P< 0.05, **P< 0.01, ***P< 0.001. Abbreviations: CD2AP, CD2-associated protein; CBS, Cystathionine β-synthase; CSE, Cystathionine γ-lyase; PI3K, Phosphoinositide 3-Kinase; AKT, Protein Kinase B.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Cardiac complications occur in MsrB2 KO DM. A . Glucose tolerance test (GTT) in WT and MsrB2 KO under nonDM and DM conditions. B . Body weight in WT and MsrB2 KO DM mice during HFD feeding. The body weight was weighed every week. C . Electron microscopy (EM) images in WT and MsrB2 KO DM mice. The white arrow indicates the mitochondria (autophagosome and autolysosome). The black triangle indicates the membrane structure. A white triangle indicates an abnormal Z-line. D . H&E and trichrome staining of histological sections of the heart of WT and MsrB2 KO mice under nonDM and DM MsrB2 conditions. E . Fibrotic areas were quantified using the ImageJ (FIJI) analyzer on histological sections. The two-way ANOVA analysis was performed for p values

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Figure Lengend Snippet: Cardiac complications occur in MsrB2 KO DM. A . Glucose tolerance test (GTT) in WT and MsrB2 KO under nonDM and DM conditions. B . Body weight in WT and MsrB2 KO DM mice during HFD feeding. The body weight was weighed every week. C . Electron microscopy (EM) images in WT and MsrB2 KO DM mice. The white arrow indicates the mitochondria (autophagosome and autolysosome). The black triangle indicates the membrane structure. A white triangle indicates an abnormal Z-line. D . H&E and trichrome staining of histological sections of the heart of WT and MsrB2 KO mice under nonDM and DM MsrB2 conditions. E . Fibrotic areas were quantified using the ImageJ (FIJI) analyzer on histological sections. The two-way ANOVA analysis was performed for p values

Article Snippet: According to the manufacturer’s protocol, MsrB2-GFP, RFP-Pakin, and Cherry LC3 were transfected into H9C2 with Lipofectamine 3000 (Invitrogen, USA).

Techniques: Electron Microscopy, Membrane, Staining

Decreased autophagy and SERCA2a-PLN function in MsrB2 KO DM mice. A . A Western blot analysis of MsrB2, LC3I/II, Parkin, SERCA2a, pPNL, PNL, and GAPDH of nonDM (WT #1–3, MsrB2 KO #1–3) and DM mouse (WT #1–3, MsrB2 KO #1–6) hearts. GAPDH served as a loading control. B – D . Quantification of MsrB2, LC3I/II, Parkin, SERCA2a, pPNL/PNL, and GAPDH signal intensity. E . Quantitative RT-PCR analysis MsrB2 and SERCA2a transcript levels in nonDM (WT #1–5, MsrB2 KO #1–5) and DM (WT #1–3, MsrB2 KO #1–6) mouse hearts. The two-way ANOVA analysis was performed for p values

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Figure Lengend Snippet: Decreased autophagy and SERCA2a-PLN function in MsrB2 KO DM mice. A . A Western blot analysis of MsrB2, LC3I/II, Parkin, SERCA2a, pPNL, PNL, and GAPDH of nonDM (WT #1–3, MsrB2 KO #1–3) and DM mouse (WT #1–3, MsrB2 KO #1–6) hearts. GAPDH served as a loading control. B – D . Quantification of MsrB2, LC3I/II, Parkin, SERCA2a, pPNL/PNL, and GAPDH signal intensity. E . Quantitative RT-PCR analysis MsrB2 and SERCA2a transcript levels in nonDM (WT #1–5, MsrB2 KO #1–5) and DM (WT #1–3, MsrB2 KO #1–6) mouse hearts. The two-way ANOVA analysis was performed for p values

Article Snippet: According to the manufacturer’s protocol, MsrB2-GFP, RFP-Pakin, and Cherry LC3 were transfected into H9C2 with Lipofectamine 3000 (Invitrogen, USA).

Techniques: Western Blot, Control, Quantitative RT-PCR

ROS induced methionine sulfoxidation (MetO) and mitochondrial dysfunction in the heart of diabetic mice. A . Blood Glucose in nonDM and DM mice. B . Western blot analysis of MetO and MsrB2 in nonDM (#1–3) and DM mouse hearts (DM #1–3). GAPDH served as a loading control. C . Quantification of MetO and MsrB2 signal intensity. D . Western blot analysis of Parkin and LC3II in nonDM (#1–3) and DM mouse hearts (DM #1–3). GAPDH served as a loading control. E . Quantification of Parkin and LC3II signal intensity. F . Western blot analysis of MsrB2 and LC3I/II in nonDM (#1–3), DM mouse hearts (DM #1–3), and DM with NAC treatment (#1–5). GAPDH served as a loading control. G . Quantification of MsrB2 and LC3II signal intensity. H . Electron microscopy (EM) in nonDM mice. The white body indicates a mitophagy structure. I . EM in DM mice. The White body indicates mitophagy (autophagosome and autolysosome) structure. J . Quantification of mitochondria size in nonDM and DM mice. K . Western blot analysis of DRP1 and DRP1 and Mfn2 in nonDM (#1–3) and DM mouse hearts (DM #1–3). GAPDH served as a loading control. L . Quantification of DRP1signal intensity. The nonparametric t -test was performed for p values

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Figure Lengend Snippet: ROS induced methionine sulfoxidation (MetO) and mitochondrial dysfunction in the heart of diabetic mice. A . Blood Glucose in nonDM and DM mice. B . Western blot analysis of MetO and MsrB2 in nonDM (#1–3) and DM mouse hearts (DM #1–3). GAPDH served as a loading control. C . Quantification of MetO and MsrB2 signal intensity. D . Western blot analysis of Parkin and LC3II in nonDM (#1–3) and DM mouse hearts (DM #1–3). GAPDH served as a loading control. E . Quantification of Parkin and LC3II signal intensity. F . Western blot analysis of MsrB2 and LC3I/II in nonDM (#1–3), DM mouse hearts (DM #1–3), and DM with NAC treatment (#1–5). GAPDH served as a loading control. G . Quantification of MsrB2 and LC3II signal intensity. H . Electron microscopy (EM) in nonDM mice. The white body indicates a mitophagy structure. I . EM in DM mice. The White body indicates mitophagy (autophagosome and autolysosome) structure. J . Quantification of mitochondria size in nonDM and DM mice. K . Western blot analysis of DRP1 and DRP1 and Mfn2 in nonDM (#1–3) and DM mouse hearts (DM #1–3). GAPDH served as a loading control. L . Quantification of DRP1signal intensity. The nonparametric t -test was performed for p values

Article Snippet: According to the manufacturer’s protocol, MsrB2-GFP, RFP-Pakin, and Cherry LC3 were transfected into H9C2 with Lipofectamine 3000 (Invitrogen, USA).

Techniques: Western Blot, Control, Electron Microscopy

Electrophysiological dysfunctions occurred in MsrB2 KO DM mice. A ~ I . ECG recording results in nonDM (WT #1–4, MsrB2 KO #1–5) and DM (WT #1–11, MsrB2. KO #1–9) mouse hearts

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Figure Lengend Snippet: Electrophysiological dysfunctions occurred in MsrB2 KO DM mice. A ~ I . ECG recording results in nonDM (WT #1–4, MsrB2 KO #1–5) and DM (WT #1–11, MsrB2. KO #1–9) mouse hearts

Article Snippet: According to the manufacturer’s protocol, MsrB2-GFP, RFP-Pakin, and Cherry LC3 were transfected into H9C2 with Lipofectamine 3000 (Invitrogen, USA).

Techniques:

MsrB2 induces autophagy by LC3 activation. A . Western blot analysis of MsrB2 and LC3I/II in H9C2 after MsrB2-GFP transfection. MsrB2 induced LC3II in a concentration-dependent manner. GAPDH served as a loading control. B . Quantification of MsrB2 and LC3II signal intensity. C . Western blot analysis of MsrB2, SERCA2a, and bMHC in WT and MsrB2 KO NMCM. After 4 h of starvation, 5.5 or 25 mM glucose was treated for 48 h. GAPDH served as a loading control. D . Quantification SERCA2a and bMHC signal intensity. E . Western blot analysis of MsrB2, LC3I/II, pDRP1, and DRP1 in NMCM after Ad-MsrB2-GFP infection. Ad-MsrB2-GFP 50 m.o.i infected into NMCM. Then, 5.5 or 25 mM glucose was treated for 48 h after 4 h starvation. GAPDH served as a loading control. F . Quantification of LC3II and pDRP1 signal intensity. The one-way ANOVA analysis was performed for p values

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Figure Lengend Snippet: MsrB2 induces autophagy by LC3 activation. A . Western blot analysis of MsrB2 and LC3I/II in H9C2 after MsrB2-GFP transfection. MsrB2 induced LC3II in a concentration-dependent manner. GAPDH served as a loading control. B . Quantification of MsrB2 and LC3II signal intensity. C . Western blot analysis of MsrB2, SERCA2a, and bMHC in WT and MsrB2 KO NMCM. After 4 h of starvation, 5.5 or 25 mM glucose was treated for 48 h. GAPDH served as a loading control. D . Quantification SERCA2a and bMHC signal intensity. E . Western blot analysis of MsrB2, LC3I/II, pDRP1, and DRP1 in NMCM after Ad-MsrB2-GFP infection. Ad-MsrB2-GFP 50 m.o.i infected into NMCM. Then, 5.5 or 25 mM glucose was treated for 48 h after 4 h starvation. GAPDH served as a loading control. F . Quantification of LC3II and pDRP1 signal intensity. The one-way ANOVA analysis was performed for p values

Article Snippet: According to the manufacturer’s protocol, MsrB2-GFP, RFP-Pakin, and Cherry LC3 were transfected into H9C2 with Lipofectamine 3000 (Invitrogen, USA).

Techniques: Activation Assay, Western Blot, Transfection, Concentration Assay, Control, Infection

ROS induced methionine sulfoxidation (MetO) in human diabetic hearts. A . Western blot analysis of MetO in normal (NH #1–3) and diabetic human heart tissue(DH #1–6). GAPDH served as a loading control. B . Quantification of MetO signal intensity. C . Western blot analysis of methionine sulfoxide reductase A and B2 (MsrA and B2) in normal (NH #1–3) and diabetic heart tissue (DH #1–6). GAPDH served as loading a loading control. D . Quantification of MsrA and MsrB2 signal intensity. E . Western blot analysis of LC3l/II and p62 in normal (NH #1–3) and diabetic heart tissue (DH #1–6). GAPDH served as a loading control. F . Quantification of LC3l/II and p62 signal intensity. G . Western blot analysis of DRP1 and OPA1 in normal (NH #1–3) and diabetic heart tissues (DH #1–6). GAPDH served as a loading control. H . Quantification of MetO intensity

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Figure Lengend Snippet: ROS induced methionine sulfoxidation (MetO) in human diabetic hearts. A . Western blot analysis of MetO in normal (NH #1–3) and diabetic human heart tissue(DH #1–6). GAPDH served as a loading control. B . Quantification of MetO signal intensity. C . Western blot analysis of methionine sulfoxide reductase A and B2 (MsrA and B2) in normal (NH #1–3) and diabetic heart tissue (DH #1–6). GAPDH served as loading a loading control. D . Quantification of MsrA and MsrB2 signal intensity. E . Western blot analysis of LC3l/II and p62 in normal (NH #1–3) and diabetic heart tissue (DH #1–6). GAPDH served as a loading control. F . Quantification of LC3l/II and p62 signal intensity. G . Western blot analysis of DRP1 and OPA1 in normal (NH #1–3) and diabetic heart tissues (DH #1–6). GAPDH served as a loading control. H . Quantification of MetO intensity

Article Snippet: According to the manufacturer’s protocol, MsrB2-GFP, RFP-Pakin, and Cherry LC3 were transfected into H9C2 with Lipofectamine 3000 (Invitrogen, USA).

Techniques: Western Blot, Control

Summary. Increased MsrB2 expression in diabetic mouse hearts suppressed cardiac complications. These results were confirmed by inducing diabetes in mice with suppressed MsrB2 expression (MsrB2 KO). In MsrB2 KO DM, the expression of SERCA2a, which regulates myocardial contractility, was reduced, cardiac fibrosis increased, and the number of mitochondrial structural abnormalities increased

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Figure Lengend Snippet: Summary. Increased MsrB2 expression in diabetic mouse hearts suppressed cardiac complications. These results were confirmed by inducing diabetes in mice with suppressed MsrB2 expression (MsrB2 KO). In MsrB2 KO DM, the expression of SERCA2a, which regulates myocardial contractility, was reduced, cardiac fibrosis increased, and the number of mitochondrial structural abnormalities increased

Article Snippet: According to the manufacturer’s protocol, MsrB2-GFP, RFP-Pakin, and Cherry LC3 were transfected into H9C2 with Lipofectamine 3000 (Invitrogen, USA).

Techniques: Expressing

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: Mouse heart tissue lysates and cell lysates (after transient transfection) were mixed with the specific target antibody [1 μg of LC3 anti-rabbit antibody (Abcam) and 2 μg of Parkin anti-goat antibody (Abcam) or 1.5 μg of Parkin anti-rabbit antibody (Abcam) for Parkin IP; 1 μg of MsrB2 anti-rabbit antibody (Yale) for MsrB2 IP; and GFP-Trap bead (Chromotek, USA), and the same species IgG control with HC] and incubated overnight at 4 °C.

Techniques: Electron Microscopy, Membrane, Staining

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Western Blot, Control, Quantitative RT-PCR

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Western Blot, Control, Electron Microscopy

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques:

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Activation Assay, Western Blot, Transfection, Concentration Assay, Control, Infection

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Western Blot, Control

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Expressing

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: We purchased Parkin and MsrB2 ORF clones from OriGene (USA) and subcloned them into RFP- and GFP-tagged vectors, respectively.

Techniques: Electron Microscopy, Membrane, Staining

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: We purchased Parkin and MsrB2 ORF clones from OriGene (USA) and subcloned them into RFP- and GFP-tagged vectors, respectively.

Techniques: Western Blot, Control, Quantitative RT-PCR

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: We purchased Parkin and MsrB2 ORF clones from OriGene (USA) and subcloned them into RFP- and GFP-tagged vectors, respectively.

Techniques: Western Blot, Control, Electron Microscopy

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: We purchased Parkin and MsrB2 ORF clones from OriGene (USA) and subcloned them into RFP- and GFP-tagged vectors, respectively.

Techniques:

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: We purchased Parkin and MsrB2 ORF clones from OriGene (USA) and subcloned them into RFP- and GFP-tagged vectors, respectively.

Techniques: Activation Assay, Western Blot, Transfection, Concentration Assay, Control, Infection

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: We purchased Parkin and MsrB2 ORF clones from OriGene (USA) and subcloned them into RFP- and GFP-tagged vectors, respectively.

Techniques: Western Blot, Control

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Article Snippet: We purchased Parkin and MsrB2 ORF clones from OriGene (USA) and subcloned them into RFP- and GFP-tagged vectors, respectively.

Techniques: Expressing

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Electron Microscopy, Membrane, Staining

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Western Blot, Control, Quantitative RT-PCR

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Western Blot, Control, Electron Microscopy

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques:

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Activation Assay, Western Blot, Transfection, Concentration Assay, Control, Infection

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Western Blot, Control

Journal: Diabetology & Metabolic Syndrome

Article Title: Methionine sulfoxide reductase B2 protects against cardiac complications in diabetes mellitus

doi: 10.1186/s13098-024-01390-0

Techniques: Expressing

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