Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA expression is upregulated in CAVD. A H&E staining showing the normal and calcific human aortic valve tissues and immunohistochemical staining showing P21 and MSRA expression in the normal and calcific human aortic valve tissues ( n = 6 for each group), scale bar = 500 μm. B Western blot analysis and quantification of OCN, RUNX2, OPN, and MSRA protein expression in calcified VICs and controls ( n = 6 for each group). C Immunofluorescence staining and quantification of RUNX2 (red) and MSRA (green) in calcified VICs and controls ( n = 6 for each group), scale bar = 20 μm. D Flow cytometry analysis and the mean fluorescence intensity of RUNX2 and MSRA in calcified VICs and controls ( n = 4 for each group). Data are presented as mean ± SEM and compared by Student’s t -test. MSRA, methionine sulfoxide reductase A; CAVD, calcific aortic valve disease; H&E, hematoxylin-and eosin; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Expressing, Staining, Immunohistochemical staining, Western Blot, Immunofluorescence, Flow Cytometry, Fluorescence

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates VIC senescence in vitro. A Western blot analysis and quantification of P21, OCN, RUNX2, OPN, and MSRA protein expression in VICs under H 2 O 2 stimulus after MSRA silencing ( n = 6 for each group). B Immunofluorescence staining and quantification of P21 (magenta) and RUNX2 (red) in VICs under H 2 O 2 stimulus after MSRA silencing ( n = 6 for each group), scale bar = 20 μm. C Western blot analysis and quantification of P21, OCN, RUNX2, OPN, and MSRA protein expression in VICs under H 2 O 2 stimulus after MSRA overexpression ( n = 6 for each group). D Immunofluorescence staining and quantification of P21 (magenta) and RUNX2 (red) in VICs under H 2 O 2 stimulus after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; P21, cyclin-dependent kinase inhibitor 1 A; H 2 O 2 , hydrogen peroxide; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: In Vitro, Western Blot, Expressing, Immunofluorescence, Staining, Over Expression

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA inhibits the TLR2/NF-κB pathway during the osteogenic differentiation of VICs. A Volcano plot representing differentially expressed proteins in VICs treated with siRNAMSRA versus siRNAscramble under osteogenic medium conditions. Differentially expressed proteins were screened upon the threshold of adjusted P -value < 0.05 and |log2 (fold change)|≥ 1. Upregulated proteins are presented in red dots and downregulated proteins are presented in blue dots ( n = 3 for each group). B The bubble plot illustrating KEGG pathway enrichment analysis of differentially expressed proteins between depleted MSRA VICs and control VICs under osteogenic medium induction ( n = 3 for each group). The Y -axis represents the top 20 different KEGG terms, the X -axis represents protein rich factor enriched in relative KEGG terms, the circle size refers to protein numbers, and the color represents P -value. C Western blot analysis and quantification of TLR2, pNF-κB p65/total NF-κB p65 expression following MSRA silencing ( n = 6 for each group). D Immunofluorescence staining and quantification of TLR2 (yellow) in VICs during the calcification process following MSRA silencing, scale bar = 20 μm. E Immunofluorescence staining and quantification of NF-κB (green) following MSRA silencing ( n = 6 for each group), scale bar = 20 μm. F Western blot analysis and quantification of TLR2, pNF-κB p65/total NF-κB p65 expression following MSRA overexpression ( n = 6 for each group). G Immunofluorescence staining and quantification of TLR2 (yellow) in VICs during the calcification process following MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. H Immunofluorescence staining and quantification of NF-κB (green) following MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; TLR2, toll-like receptor 2; pNF-κB, phosphorylated nuclear factor-κB; KEGG, Kyoto Encyclopedia of Genes and Genomes; VICs, valvular interstitial cells; OM, osteogenic medium; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Control, Western Blot, Expressing, Immunofluorescence, Staining, Over Expression

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates osteogenic differentiation of VICs by inhibiting the TLR2/NF-κB pathway. A Western blot analysis and quantification of OCN, RUNX2, and OPN protein expression when VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 (0.1 μg/mL) after MSRA overexpression ( n = 6 for each group). B Immunofluorescence staining and quantification of RUNX2 (red) when human VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. C Flow cytometry analysis and the mean fluorescence intensity of RUNX2 when human VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 4 for each group). D Alkaline phosphatase staining when VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 6 for each group), scale bar = 5 mm and 500 μm. Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; VICs, valvular interstitial cells; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Western Blot, Expressing, Cell Culture, Over Expression, Immunofluorescence, Staining, Flow Cytometry, Fluorescence

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA overexpression alleviates aortic valve calcification and senescence in vivo. A Echocardiographic data peak transvalvular jet velocity in ApoE −/− mice ( n = 9 for each ND group, n = 10 for each HCD group). B H&E staining of the aortic valve leaflets in ApoE −/− mice. C Von Kossa staining of the aortic valve leaflets in ApoE −/− mice ( n = 9 for each ND group, n = 10 for each HCD group), scale bar = 250 μm. D Immunohistochemical staining showed P21 in ApoE −/− mice ( n = 9 for each ND group, n = 10 for each HCD group), scale bar = 250 μm. E The inflammatory cytokines in ApoE. −/− mice ( n = 9 for each ND group, n = 10 for each HCD group). Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; H&E, hematoxylin-and eosin; AAV2, adeno-associated virus subtype 2; ND, normal diet; HCD, high cholesterol diet; P21, cyclin-dependent kinase inhibitor 1 A; MCP-1, monocyte chemoattractant protein-1; IL, interleukin; TNF, tumor necrosis factor. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Over Expression, In Vivo, Staining, Immunohistochemical staining, Virus

Journal: Cell reports

Article Title: Inhibiting G6PD by quercetin promotes degradation of EGFR T790M mutation.

doi: 10.1016/j.celrep.2023.113417

Figure Lengend Snippet: Figure 3. Quercetin induces M790 oxidation for EGFRT790M degradation through impairing NADPH-dependent MsrA activity (A) FLAG-tagged EGFRT790M immunoprecipitated from HEK293T cells was analyzed by Matrix-Assisted Laser Desorption Ionization Tandem Time-of-Flight Mass Spectrometry (MALDI-TOF/TOF MS). M790 residue was identified to be oxidized. See also Table S3. (B) H820 and H1975 cells were transfected with or without G6PD siRNA for 48 h. (C) H820 and H1975 cells overexpressing escalating FLAG-G6PD were treated with or without 20 mM Que for 48 h. (D) H820 and H1975 cells overexpressed with or without FLAG-ME1 or FLAG-IDH1 were treated with or without 20 mM Que for 48 h. (E) H820 and H1975 cells were treated with or without 20 mM Que or 20 mM NADPH supplementation for 48 h. (F) H820 and H1975 cells with or without MsrA depletion were treated with or without 20 mM Que for 48 h. (G) H820 and H1975 cells with or without MsrA depletion were treated with or without 20 mM NADPH supplementation for 24 h. (H–J) H820 and H1975 cells incubated with or without Ox4 were treated with or without 20 mM Que for 24 h (H) or depleted with or without siRNAs targeting G6PD (I) or MsrA (J). ReACT assay was performed to detect the levels of methionine oxidation in EGFR. (B–H) Immunoprecipitation and immunoblotting analyses were performed with the indicated antibodies. See also Figure S4.

Article Snippet: G6PD siRNA, ME1 siRNA and MsrA siRNA were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activity Assay, Immunoprecipitation, Mass Spectrometry, Residue, Transfection, Incubation, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: The Interaction of Selective A1 and A2A Adenosine Receptor Antagonists with Magnesium and Zinc Ions in Mice: Behavioural, Biochemical and Molecular Studies

doi: 10.3390/ijms22041840

Figure Lengend Snippet: Effect of co-administration of DPCPX and istradefylline with Mg 2+ and Zn 2+ on gene expression ( A ) Adora1 , ( B ) Slc6a15 , ( C ) Comt , ( D ) Ogg1 , ( E ) MsrA , and ( F ) Nrf2 in mice prefrontal cortex. DPCPX, Mg 2+ and saline were administered i.p. 30 min, whereas istradefylline p.o. and Zn 2+ i.p. 60 min prior decapitation. The data are presented as the means ± SEM. Each experimental group consisted of 10 animals. ( A ) * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. NaCl-treated group; ^^^^ p < 0.0001 vs. DPCPX-treated group; ### p < 0.001 vs. istradefylline-treated group; ++++ p < 0.0001 vs. Mg 2+ -treated group; &&&& p < 0.0001 vs. Zn 2+ -treated group (two-way ANOVA followed by Bonferroni’s post hoc test). ( B ) * p < 0.05, **** p < 0.0001 vs. NaCl-treated group; #### p < 0.0001 vs. istradefylline-treated group; &&&& p < 0.0001 vs. Zn 2+ -treated group (two-way ANOVA followed by Bonferroni’s post hoc test). ( C ) * p < 0.05, ** p < 0.01, **** p < 0.0001 vs. NaCl-treated group; ^^^ p < 0.001, ^^^^ p < 0.0001 vs. DPCPX-treated group; # p < 0.05, ## p < 0.01 vs. istradefylline-treated group; ++++ p < 0.0001 vs. Mg 2+ -treated group; &&& p < 0.001, &&&& p < 0.0001 vs. Zn 2+ -treated group (two-way ANOVA followed by Bonferroni’s post hoc test). ( D ) *** p < 0.001, **** p < 0.0001 vs. NaCl-treated group; ^^^^ p < 0.0001 vs. DPCPX-treated group; # p < 0.05, ### p < 0.001 vs. istradefylline-treated group; + p < 0.05 vs. Mg 2+ -treated group (two-way ANOVA followed by Bonferroni’s post hoc test). ( E ) * p < 0.05, ** p < 0.01 vs. NaCl-treated group; #### p < 0.0001 vs. istradefylline-treated group; + p < 0.05, +++ p < 0.001 vs. Mg 2+ -treated group; && p < 0.01 vs. Zn 2+ -treated group (two-way ANOVA followed by Bonferroni’s post hoc test). ( F ) *** p < 0.001, **** p < 0.0001 vs. NaCl-treated group; ^^^^ p < 0.0001 vs. DPCPX- treated group; # p < 0.05 vs. istradefylline-treated group; ++++ p < 0.0001 vs. Mg 2+ -treated group; &&& p < 0.001, &&&& p < 0.0001 vs. Zn 2+ -treated group (two-way ANOVA followed by Bonferroni’s post hoc test).

Article Snippet: Msra , Methionine sulfoxide reductase A , NM_001253712.1 NM_001253714.1 NM_001253716.1 NM_026322.4 , Mm00452737_m1 , 69.

Techniques: Gene Expression, Saline

Journal: International Journal of Molecular Sciences

Article Title: The Interaction of Selective A1 and A2A Adenosine Receptor Antagonists with Magnesium and Zinc Ions in Mice: Behavioural, Biochemical and Molecular Studies

doi: 10.3390/ijms22041840

Figure Lengend Snippet: Gene symbols, gene names, GenBank reference sequence accession numbers, assay IDs and amplicon lengths (bp).

Article Snippet: Msra , Methionine sulfoxide reductase A , NM_001253712.1 NM_001253714.1 NM_001253716.1 NM_026322.4 , Mm00452737_m1 , 69.

Techniques: Sequencing, Amplification, Derivative Assay, Binding Assay

Journal: Lipids in Health and Disease

Article Title: Tetradecylthiopropionic acid induces hepatic mitochondrial dysfunction and steatosis, accompanied by increased plasma homocysteine in mice

doi: 10.1186/s12944-016-0192-9

Figure Lengend Snippet: Hepatic gene expression levels a

Article Snippet: Real-time PCR was performed with Sarstedt 384 well multiply-PCR Plates (Sarstedt Inc., Newton, NC, USA) on the following genes, using probes and primers from Applied Biosystems: Chka (Mm00442759_m1), Chkb (Mm04213225_s1), Chdh (Mm00549261_m1), Bhmt (Mm04210521_g1), Dmgdh (Mm00512022_m1), Sardh (Mm00454657_m1), Shmt1 (Mm00486110_m1), Shmt2 (Mm00659512_g1), Gnmt (Mm00494688_m1), Mtr (Mm01340053_m1), Mtrr _(Mm00549978_m1), Msra (Mm00452738_m1), Msrb2 (Mm00512937_m1), Cbs (Mm00460654_m1), Cth (Mm00461247_m1), Ppargc1a (Mm00447183), Pparg (Mm00440945), Ppara (Mm00440939), CD36 / Fat (Mm00432403), Fabp1 (Mm00444340), Lipe (Mm00495359), Acly (Mm01302282_m1), Slc25a20 (Mm00451571_m1), Lipc (Mm00433975), Apob (Mm01545156_m1), IL1β (Mm01336189_m1), Sod1 (Mm01344233_m1), Sod2 (Mm01313000_m1), Tnfα (Mm00443258_m1) and Pcyt1a (Mm00447774_m1).

Techniques: Gene Expression, Control, Binding Assay, Antioxidant Activity Assay