Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Primer used for RT-pPCR.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Act1 is expressed in healthy and periodontitis gingival tissue. (A) Representative immunohistochemistry images of healthy/periodontitis gingival tissue (human and mice) show Act1 expression. Quantitative analysis of Act1 positive cells in healthy and periodontitis human gingival tissue (B) , and mice periodontal tissue (C) . Data are presented as mean ± SD ( n = 6). WT, wildtype.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: Immunohistochemistry, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Periodontitis and alveolar bone loss were aggravated in anti-Act1 mice compared to wildtype. (A) Micro-CT images of the healthy and periodontitis-affected maxilla of wildtype and anti-Act1 mice. Quantitative analysis of CEJ-ABC distance (B) , BMD (C) from micro-CT images. (D) H&E stained histological images of PAPT of wild-type and anti-Act1 mice. (E) Quantitative analysis of CEJ-ABC distance from histological images. (F) TRAP stained histological images of PAPT of wild-type and anti-Act1 mice. (G) Quantitative analysis of osteoclasts in PAPT. Data are presented as mean ± SD ( n = 6). The significant difference between the groups, *** P < 0.001. CEJ, cement-enamel junction; ABC, alveolar bone crest; Red dot line, CEJ level; Black dot line, ABC level; blue line, CEJ-ABC distance; yellow arrow, osteoclasts; WT, wild-type; and BMD, bone mineral density.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: Micro-CT, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Macrophage infiltration in PAPT was enhanced in anti-Act1 mice compared to wildtype. (A) Immunohistochemistry images of PAPT. Quantitative analysis of CD34 expressing MVD (B) , CD45 expressing pan leucocytes (C) , and F4/80 expressing macrophages (D) , n = 6. (E) FACs images of macrophages sorting using FITC-CD11b and PE-F4/80. (F) Quantitative FACs data. Calculated the ratio of CD11b + F4/80+ cells, n = 8. The significant difference between the groups, ** P < 0.01, ns, not significant. MVD, Microvessel density; WT, wildtype.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: Immunohistochemistry, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: (A–F) mRNA level expression of inflammatory cytokines and macrophage migration-related factors in PAPT of wildtype and anti-Act1 mice. (G–I) Protein level expression of inflammatory cytokines in PAPT of wildtype and anti-Act1 mice. Data are presented as mean ± SD ( n = 6). The significant difference between the groups, ** P < 0.01, *** P < 0.001. WT, wildtype.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: Expressing, Migration

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Lipopolysaccharide -treated peritoneal macrophages from wildtype and anti-Act1 mice showed a differential expression of 2190 mRNAs. (A) Heat map, (B) KEGG pathway analysis, and (C) Volcano map showing differentially expressed mRNAs. Red color intensity indicates upregulation, and blue color intensity indicates the downregulation of mRNA expression.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Lipopolysaccharide -treated peritoneal macrophages from anti-Act1 mice differentially upregulate the mRNA expression of TNF-NF-κB signaling pathway-related proteins. (A) Heat map, and (B) bar figures showing differentially upregulated mRNAs. (C–E) Protein level expressions of inflammatory cytokines in LPS-treated peritoneal macrophages from wildtype and anti-Act1 mice (measured by ELISA), n = 6. (F) Representative western bolt images of p65 and P-p65 expression. (G) Quantification of western blots ( n = 3). Data are presented as mean ± SD ( n = 6). The significant difference between the groups, ** P < 0.01, *** P < 0.001. WT, wildtype. Red color intensity indicates upregulation, and blue color intensity indicates the downregulation of gene expression.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Lipopolysaccharide -treated peritoneal macrophages from anti-Act1 mice showed higher migration and M1-polarization. (A) Representative images of macrophage migration. (B) Quantitative analysis of macrophage migration, n = 3. (C) CCL2 protein expression (ELISA, n = 6). Heat map (D) , and bar figures of differentially expressed macrophage M1-polarization-related genes (E) . (F) CD16/32 protein level expression analyzed by ELISA, n = 6. (G) FACs images of mice periodontal macrophages sorting using CD16/32 specific antibody. (H) Quantitative FACs data, n = 6. The significant difference between the groups, ** P < 0.01, *** P < 0.001. WT, wildtype. Red color intensity indicates upregulation, and blue color intensity indicates the downregulation of gene expression.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: Migration, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Downregulation of Macrophage-Specific Act-1 Intensifies Periodontitis and Alveolar Bone Loss Possibly via TNF/NF-κB Signaling

doi: 10.3389/fcell.2021.628139

Figure Lengend Snippet: Effect of macrophage-specific downregulation of Act1 on the pathophysiology of mice periodontitis.

Article Snippet: Tissue sections were then blocked with 10% bovine serum albumin (BSA) and incubated with primary antibodies: anti-Act1 (bs6202R, BIOSS, China), anti-CD34 (ab81289, Abcam, Cambridge, United Kingdom), anti-CD45 (20103-1-AP, Proteintech, United States), anti-F4/80(70076, CST, United States) or isotope controls for overnight at 4°C.

Techniques: