Review

pmscv pig mir497 mir 195 (Addgene inc)

Addgene inc is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Addgene inc pmscv pig mir497 mir 195
Pmscv Pig Mir497 Mir 195, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pmscv pig mir497 mir 195/product/Addgene inc
Average 91 stars, based on 1 article reviews

pmscv pig mir497 mir 195 - by Bioz Stars, 2026-04

91/100 stars

Images

Image Search Results

miR-146a –/– mice (12 wk of age) displayed elevated systemic and gut IgA + B cells, IgA-secreting cells, and kidney IgA deposition, as hallmark of IgA nephropathy. (A) IgA + B cells in spleen, bone marrow, mesenteric lymph nodes (MLNs) and Peyer’s patches as analyzed by flow cytometry. (B) Fecal bacteria-bound IgA as analyzed by flow cytometry. (C) IgM + and IgA + B cells in the lamina propria and MLNs of miR-146a and miR-146a +/+ mice as visualized by fluorescence microscopy. (D) ELISPOT analysis of IgM-, IgG1- and IgA-secreting cells (ASCs) in spleen and MLNs. (E) Photomicrographs of kidney sections from miR-146a –/– and miR-146a +/+ mice, after fluorescence staining for mouse IgA (left) or hematoxylin and eosin (H&E) staining (right). Data are from one representative of three independent experiments yielding similar results; or mean ± SEM of 3-4 miR-146a –/– or miR-146a +/+ mice from three independent experiments ( D , right panel). ** p < 0.01, ns, not significant, unpaired t -test.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: miR-146a –/– mice (12 wk of age) displayed elevated systemic and gut IgA + B cells, IgA-secreting cells, and kidney IgA deposition, as hallmark of IgA nephropathy. (A) IgA + B cells in spleen, bone marrow, mesenteric lymph nodes (MLNs) and Peyer’s patches as analyzed by flow cytometry. (B) Fecal bacteria-bound IgA as analyzed by flow cytometry. (C) IgM + and IgA + B cells in the lamina propria and MLNs of miR-146a and miR-146a +/+ mice as visualized by fluorescence microscopy. (D) ELISPOT analysis of IgM-, IgG1- and IgA-secreting cells (ASCs) in spleen and MLNs. (E) Photomicrographs of kidney sections from miR-146a –/– and miR-146a +/+ mice, after fluorescence staining for mouse IgA (left) or hematoxylin and eosin (H&E) staining (right). Data are from one representative of three independent experiments yielding similar results; or mean ± SEM of 3-4 miR-146a –/– or miR-146a +/+ mice from three independent experiments ( D , right panel). ** p < 0.01, ns, not significant, unpaired t -test.

Article Snippet: The miR-146a expression retroviral construct pMSCV-PIG-miR-146a ( ) was a gift from Joshua Mendell (Addgene plasmid # 64234) and the pMSCV-PIG (Puro IRES GFP) empty vector was a gift from David Bartel (Addgene plasmid # 21654).

Techniques: Flow Cytometry, Bacteria, Fluorescence, Microscopy, Enzyme-linked Immunospot, Staining

Increased IgA production in miR-146a –/– mice. (A) miR-146a –/– and miR-146a +/+ mice were administered OVA together with CT via intragastric gavage once a week for 3 wks and sacrificed one wk after the last OVA administration for analysis of antibodies, B cells and ASCs. Total (B) and OVA-binding (C) IgM, IgG1, IgA and IgE titers, as measured by ELISA in serum and feces. ** p < 0.01, * p < 0.05, ns, not significant, unpaired t -test. Data are mean ± SEM of 4-7 miR-146a –/– or miR-146a +/+ mice from three independent experiments.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: Increased IgA production in miR-146a –/– mice. (A) miR-146a –/– and miR-146a +/+ mice were administered OVA together with CT via intragastric gavage once a week for 3 wks and sacrificed one wk after the last OVA administration for analysis of antibodies, B cells and ASCs. Total (B) and OVA-binding (C) IgM, IgG1, IgA and IgE titers, as measured by ELISA in serum and feces. ** p < 0.01, * p < 0.05, ns, not significant, unpaired t -test. Data are mean ± SEM of 4-7 miR-146a –/– or miR-146a +/+ mice from three independent experiments.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Increased IgA production, IgA + B cells and IgA-ASCs in μMT/miR-146a –/– chimeric mice. μMT/miR-146a –/– and μMT/miR-146a +/+ chimeric mice were constructed by mixed bone marrow adaptive transfer. (A) All mice were administered OVA together with CT via intragastric gavage once a week for 3 wks and sacrificed one wk after the last OVA administration for analysis of antibodies, B cells and ASCs. Total and OVA-binding IgM, IgG1, IgA and IgE titers in sera (B) and feces (C) as analyzed by ELISAs. (D) Fecal bacteria-bound IgA as analyzed by flow cytometry. (E) Circulating T (CD3 + ) and B (CD19 + ) cells, Peyer’s patched IgA + B cells, IgG1 + B cells and CD138 + plasma cells, as analyzed by flow cytometry. (F) ELISPOT analysis of OVA-binding IgA ASCs in the bone marrow (BM), spleen and MLNs. *** p < 0.001, ** p < 0.01, * p < 0.05, ns, not significant, unpaired t -test. Data in (B, C) and ( F , right panel) are mean ± SEM of 3-7 μMT/miR-146a –/– or μMT/miR-146a +/+ from three independent experiments. Data in (D, E) and ( F , left panel) are from one representative of three independent experiments yielding similar results.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: Increased IgA production, IgA + B cells and IgA-ASCs in μMT/miR-146a –/– chimeric mice. μMT/miR-146a –/– and μMT/miR-146a +/+ chimeric mice were constructed by mixed bone marrow adaptive transfer. (A) All mice were administered OVA together with CT via intragastric gavage once a week for 3 wks and sacrificed one wk after the last OVA administration for analysis of antibodies, B cells and ASCs. Total and OVA-binding IgM, IgG1, IgA and IgE titers in sera (B) and feces (C) as analyzed by ELISAs. (D) Fecal bacteria-bound IgA as analyzed by flow cytometry. (E) Circulating T (CD3 + ) and B (CD19 + ) cells, Peyer’s patched IgA + B cells, IgG1 + B cells and CD138 + plasma cells, as analyzed by flow cytometry. (F) ELISPOT analysis of OVA-binding IgA ASCs in the bone marrow (BM), spleen and MLNs. *** p < 0.001, ** p < 0.01, * p < 0.05, ns, not significant, unpaired t -test. Data in (B, C) and ( F , right panel) are mean ± SEM of 3-7 μMT/miR-146a –/– or μMT/miR-146a +/+ from three independent experiments. Data in (D, E) and ( F , left panel) are from one representative of three independent experiments yielding similar results.

Techniques: Construct, Binding Assay, Bacteria, Flow Cytometry, Clinical Proteomics, Enzyme-linked Immunospot

B cell miR-146a deficiency alters gut bacterial microbiome in μMT/miR-146a –/– mice. Relative abundance of fecal bacteria families and genera (A, B) in μMT/miR-146a –/– and μMT/miR-146a +/+ chimeric mice ( n = 3 per group) as determined by high-throughput 16s rRNA gene miSeq amplicon sequencing. (C) Frequency of five relatively abundant fecal bacteria families and genera in these mice. *** p < 0.001, ** p < 0.01, ns, not significant, unpaired t -test. (D) Principal component analysis of gut bacterial composition of the fecal bacteria families and genera in μMT/miR-146a –/– and μMT/miR-146a +/+ chimeric mice. Data are from three independent experiments.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: B cell miR-146a deficiency alters gut bacterial microbiome in μMT/miR-146a –/– mice. Relative abundance of fecal bacteria families and genera (A, B) in μMT/miR-146a –/– and μMT/miR-146a +/+ chimeric mice ( n = 3 per group) as determined by high-throughput 16s rRNA gene miSeq amplicon sequencing. (C) Frequency of five relatively abundant fecal bacteria families and genera in these mice. *** p < 0.001, ** p < 0.01, ns, not significant, unpaired t -test. (D) Principal component analysis of gut bacterial composition of the fecal bacteria families and genera in μMT/miR-146a –/– and μMT/miR-146a +/+ chimeric mice. Data are from three independent experiments.

Techniques: Bacteria, High Throughput Screening Assay, Amplification, Sequencing

miR-146a ablation results in increased germline Iα-Cα transcripts and enhanced CSR to IgA . (A) miR-146a –/– and miR-146a +/+ B cells were stimulated with LPS, LPS or CD154 plus IL-4, LPS or CD154 plus IL-4, IL-5, TGF-β, RA and anti-δ mAb, and cultured for 96 h. IgG1 + , IgG3 + and IgA + B cells and CD138 + plasma cells as analyzed by flow cytometry. Data are one representative of three independent experiments yielding similar results. (B) IgG1 and IgA concentrations in culture fluids from 146a –/– and miR-146a +/+ B cells stimulated by LPS plus IL-4 or LPS plus IL-4, IL-5, TGF-β, RA and anti-δ mAb, as measured by ELISA. (C) miR-146a –/– and miR-146a +/+ B cells were stimulated by LPS or CD154 plus IL-4, IL-5, TGF-β, anti-δ mAb and RA, and cultured for 72 h. Expression of germline Iμ-Cμ , Iγ1-Cγ1 , Iα-Cα and Iϵ-Cϵ transcripts, circle Iα-Cμ and post-recombination Iμ-Cα transcripts, as well as Aicda transcripts as analyzed by qRT-PCR and normalized to β-Actin. Data are ratios to miR-146a +/+ B cells (set as 1; means ± SEM of three independent experiments). ** p < 0.01, ns, not significant, paired t -test.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: miR-146a ablation results in increased germline Iα-Cα transcripts and enhanced CSR to IgA . (A) miR-146a –/– and miR-146a +/+ B cells were stimulated with LPS, LPS or CD154 plus IL-4, LPS or CD154 plus IL-4, IL-5, TGF-β, RA and anti-δ mAb, and cultured for 96 h. IgG1 + , IgG3 + and IgA + B cells and CD138 + plasma cells as analyzed by flow cytometry. Data are one representative of three independent experiments yielding similar results. (B) IgG1 and IgA concentrations in culture fluids from 146a –/– and miR-146a +/+ B cells stimulated by LPS plus IL-4 or LPS plus IL-4, IL-5, TGF-β, RA and anti-δ mAb, as measured by ELISA. (C) miR-146a –/– and miR-146a +/+ B cells were stimulated by LPS or CD154 plus IL-4, IL-5, TGF-β, anti-δ mAb and RA, and cultured for 72 h. Expression of germline Iμ-Cμ , Iγ1-Cγ1 , Iα-Cα and Iϵ-Cϵ transcripts, circle Iα-Cμ and post-recombination Iμ-Cα transcripts, as well as Aicda transcripts as analyzed by qRT-PCR and normalized to β-Actin. Data are ratios to miR-146a +/+ B cells (set as 1; means ± SEM of three independent experiments). ** p < 0.01, ns, not significant, paired t -test.

Techniques: Cell Culture, Clinical Proteomics, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

miR-146a targets 3’UTR of Smad2, Smad3 and Smad4 mRNAs and is downregulated by the stimuli that induce CSR to IgA in human and mouse B cells. (A) Alignment of miR-146a with its target sites in the 3′ UTR of human and mouse SMAD2/Smad2 , SMAD3/Smad3 and SMAD4/Smad4 mRNAs. The complementary base pairing between miRNA-146a and seed sequences within the 3’ UTRs of SMAD2/Smad2 , SMAD3/Smad3 and SMAD4/Smad4 , indicating miR-146a ability to silence SMAD2/Smad2 , SMAD3/Smad3 and SMAD4/Smad4 transcripts. Gibbs free energy (Δ G ) of nucleic acid folding and hybridization calculated by Mfold. Watson–Crick base-pairing (“|”); wobble base-pairing (“:”). (B) Expression of miRNA-146a in mouse spleen, MLN and Peyer’s patch B cells ex vivo ; and in mouse spleen B cells stimulated for 72 h with Nil, LPS, LPS plus IL-4 and TGF-β, LPS or CD154 plus IL-4, TGF-β and RA, or human B cells stimulated for 72 h with Nil, CD154, CpG, and CD154 or CpG plus TGF-β, as analyzed by qRT-PCR. Values were normalized to expression of small nuclear/nucleolar RNAs Rnu6, Snord61, Snord68, and Snord70, and depicted as relative to the expression of miRNA-146a in spleen B cells stimulated with Nil, set as 1. (C) Expression of Smad2, Smad3 and Smad4 transcripts in mouse B cells stimulated for 72 h with Nil or LPS plus IL-4, TGF-β and RA, as analyzed by qRT-PCR and normalized to β-Actin , and depicted as relative to the expression of these genes in spleen B cells stimulated with nil, set as 1. Data are mean ± SEM from three independent experiments. *** p < 0.001, ** p < 0.01, ns, not significant, unpaired t -test.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: miR-146a targets 3’UTR of Smad2, Smad3 and Smad4 mRNAs and is downregulated by the stimuli that induce CSR to IgA in human and mouse B cells. (A) Alignment of miR-146a with its target sites in the 3′ UTR of human and mouse SMAD2/Smad2 , SMAD3/Smad3 and SMAD4/Smad4 mRNAs. The complementary base pairing between miRNA-146a and seed sequences within the 3’ UTRs of SMAD2/Smad2 , SMAD3/Smad3 and SMAD4/Smad4 , indicating miR-146a ability to silence SMAD2/Smad2 , SMAD3/Smad3 and SMAD4/Smad4 transcripts. Gibbs free energy (Δ G ) of nucleic acid folding and hybridization calculated by Mfold. Watson–Crick base-pairing (“|”); wobble base-pairing (“:”). (B) Expression of miRNA-146a in mouse spleen, MLN and Peyer’s patch B cells ex vivo ; and in mouse spleen B cells stimulated for 72 h with Nil, LPS, LPS plus IL-4 and TGF-β, LPS or CD154 plus IL-4, TGF-β and RA, or human B cells stimulated for 72 h with Nil, CD154, CpG, and CD154 or CpG plus TGF-β, as analyzed by qRT-PCR. Values were normalized to expression of small nuclear/nucleolar RNAs Rnu6, Snord61, Snord68, and Snord70, and depicted as relative to the expression of miRNA-146a in spleen B cells stimulated with Nil, set as 1. (C) Expression of Smad2, Smad3 and Smad4 transcripts in mouse B cells stimulated for 72 h with Nil or LPS plus IL-4, TGF-β and RA, as analyzed by qRT-PCR and normalized to β-Actin , and depicted as relative to the expression of these genes in spleen B cells stimulated with nil, set as 1. Data are mean ± SEM from three independent experiments. *** p < 0.001, ** p < 0.01, ns, not significant, unpaired t -test.

Techniques: Hybridization, Expressing, Ex Vivo, Quantitative RT-PCR

miR-146a deletion increases expression of Smad2, Smad3 and Smad4 , germline Iα-Cα and post-recombination Iμ-Cα transcripts. miR-146a –/– and miR-146a +/+ B cells were stimulated with LPS or CD154 plus IL-4, IL-5, TGF-β, RA and anti-δ mAb, and cultured for 72 h. (A) The expression of Smad2 , Smad3 and Smad4 as well as Iα-Cα, Iμ-Cα, Traf6, Irak1, Aicda and Prdm1 transcripts was analyzed by qRT-PCR performed in triplicate and normalized to β-Actin. (B) Expression of Smad2, Smad3 and Smad4 proteins as analyzed by immunoblotting. Densitometry quantification of immunoblotting signals was normalized to β-Actin levels and depicted as ratios of readings in miR-146a –/– and miR-146a +/+ B cells to the average value in miR-146a +/+ B cells. Data are ratios to stimulated miR-146a +/+ B cells (set as 1; means ± SEM of three independent experiments). *** p < 0.001, ** p < 0.01, * p < 0.05, ns, not significant, unpaired t -test.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: miR-146a deletion increases expression of Smad2, Smad3 and Smad4 , germline Iα-Cα and post-recombination Iμ-Cα transcripts. miR-146a –/– and miR-146a +/+ B cells were stimulated with LPS or CD154 plus IL-4, IL-5, TGF-β, RA and anti-δ mAb, and cultured for 72 h. (A) The expression of Smad2 , Smad3 and Smad4 as well as Iα-Cα, Iμ-Cα, Traf6, Irak1, Aicda and Prdm1 transcripts was analyzed by qRT-PCR performed in triplicate and normalized to β-Actin. (B) Expression of Smad2, Smad3 and Smad4 proteins as analyzed by immunoblotting. Densitometry quantification of immunoblotting signals was normalized to β-Actin levels and depicted as ratios of readings in miR-146a –/– and miR-146a +/+ B cells to the average value in miR-146a +/+ B cells. Data are ratios to stimulated miR-146a +/+ B cells (set as 1; means ± SEM of three independent experiments). *** p < 0.001, ** p < 0.01, * p < 0.05, ns, not significant, unpaired t -test.

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Western Blot

miR-146a deletion results in increased Smad2, Smad3 and Smad4 recruitment to Iα promoter. (A) Human and mouse Iα promoters share five SBEs, three of which are identical and conserved in mouse and human. (B) miR-146a –/– and miR-146a +/+ B cells were stimulated with LPS or CD154 plus IL-4, IL-5, TGF-β, anti-δ mAb and RA for 72 h. The recruitment of Smad2, Smad3 and Smad4 to the Iα promoter was analyzed by specific ChIP assays. Data are mean ± SEM from three independent experiments. ** p < 0.01, * p < 0.05, unpaired t -test. (C) Cartoon depicting the increased availability of Smad2, Smad3 and Smad4 in miR-146a –/– B cells and increased recruitment of these transcription factors to the IgH locus Iα promoter.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: miR-146a deletion results in increased Smad2, Smad3 and Smad4 recruitment to Iα promoter. (A) Human and mouse Iα promoters share five SBEs, three of which are identical and conserved in mouse and human. (B) miR-146a –/– and miR-146a +/+ B cells were stimulated with LPS or CD154 plus IL-4, IL-5, TGF-β, anti-δ mAb and RA for 72 h. The recruitment of Smad2, Smad3 and Smad4 to the Iα promoter was analyzed by specific ChIP assays. Data are mean ± SEM from three independent experiments. ** p < 0.01, * p < 0.05, unpaired t -test. (C) Cartoon depicting the increased availability of Smad2, Smad3 and Smad4 in miR-146a –/– B cells and increased recruitment of these transcription factors to the IgH locus Iα promoter.

Techniques:

Enforced expression of miR-146a in B cells reduces Smad2, Smad3 and Smad4 expression and CSR to IgA. B cells isolated from C57BL/6 mice were transduced with pMSCV-PIG-miR-146a retroviral vector expressing GFP and miR-146a or empty pMSCV-PIG retroviral vector that expression GFP, then stimulated with LPS plus IL-4, IL-5, TGF-β, anti-δ mAb and RA and cultured for 96 h. Proportions of surface IgA + B cells, and intracellular AID, Smad2, Smad3 and Smad4 levels among the retroviral vector-transduced (B220 + GFP + ) B cells were analyzed by flow cytometry. Data are from one representative of three independent experiments.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: Enforced expression of miR-146a in B cells reduces Smad2, Smad3 and Smad4 expression and CSR to IgA. B cells isolated from C57BL/6 mice were transduced with pMSCV-PIG-miR-146a retroviral vector expressing GFP and miR-146a or empty pMSCV-PIG retroviral vector that expression GFP, then stimulated with LPS plus IL-4, IL-5, TGF-β, anti-δ mAb and RA and cultured for 96 h. Proportions of surface IgA + B cells, and intracellular AID, Smad2, Smad3 and Smad4 levels among the retroviral vector-transduced (B220 + GFP + ) B cells were analyzed by flow cytometry. Data are from one representative of three independent experiments.

Techniques: Expressing, Isolation, Transduction, Retroviral, Plasmid Preparation, Cell Culture, Flow Cytometry

Epigenetic regulation of CSR to IgA by miR-146a. miR-146a targets the 3’UTRs of Smad2 , Smad3 and Smad4 mRNAs to downregulate the expression of Smad2, Smad3 and Smad4, and reduce germline Iα-Cα transcription leading to a decreased CSR to IgA and IgA production. In B cells stimulation by TGF-β, activated Smad proteins lead to miR-146 reduction, which results in enhancement of expression of Smad proteins themselves. This miR-146a/Smad loop can amplify TGF-β-induced Smads promotion of CSR to IgA.

Journal: Frontiers in Immunology

Article Title: Epigenetic Modulation of Class-Switch DNA Recombination to IgA by miR-146a Through Downregulation of Smad2, Smad3 and Smad4

doi: 10.3389/fimmu.2021.761450

Figure Lengend Snippet: Epigenetic regulation of CSR to IgA by miR-146a. miR-146a targets the 3’UTRs of Smad2 , Smad3 and Smad4 mRNAs to downregulate the expression of Smad2, Smad3 and Smad4, and reduce germline Iα-Cα transcription leading to a decreased CSR to IgA and IgA production. In B cells stimulation by TGF-β, activated Smad proteins lead to miR-146 reduction, which results in enhancement of expression of Smad proteins themselves. This miR-146a/Smad loop can amplify TGF-β-induced Smads promotion of CSR to IgA.

Techniques: Expressing

Review

Structured Review

Images

Similar Products

Image Search Results