Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 2. 6014 attenuated the enlargement of the heart size, cardiomyocytes size and prevented the accumulation of collagen. (A) Representative diagram of mice hearts under different treatment. (B) Representative images diagram of H&E staining of cardiomyocytes. (C) Representative images of Masson’s staining of total collagen in tissue sections of heart. (D) Statistical results of HW/BW. (E) Statistical results of LVW/BW. (F) Statistical areas of total collagen production versus areas of whole tissue. Data were expressed as mean±SEM from three independent experiments. **P<0.01 vs Sham. #P<0.05, ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Staining

Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 3. Effects of 6014 on the expression of hypertrophy marker proteins and fibrosis marker proteins. Protein expression of ANF (A), BNP (B), β-MHC (C), FN (D), collagen I (E) and collagen III (F) were measured by Western blot. Data were expressed as mean±SEM from duplicates in three independent experiments. **P<0.01 vs Sham. #P<0.05, ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Expressing, Marker, Western Blot

Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 4. Effects of 6014 on the expression and enzymatic activity of PARP-1. The expression of PARP-1 in nucleus (A) and whole cell (B) as well as PARylated proteins in whole cell (C). Data were expressed as mean±SEM from duplicates in three independent experiments. *P<0.05, **P<0.01 vs Sham. ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Expressing, Activity Assay

Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 6. Effects of 6014 on the regulation and activity of SIRT-1. NAD+ content analysis (A) and SIRT-1 activity (B) were analyzed. Protein expression of SIRT-1 were measured by Western blot (C). The interaction between SIRT-1 and PARP-1 (D) and the PARylation of SIRT-1 (E) were measured by co-IP assay. Data were expressed as mean±SEM from three independent experiments. *P<0.05, **P<0.01 vs Sham. #P<0.05, ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Activity Assay, Expressing, Western Blot, Co-Immunoprecipitation Assay

Journal: International Journal of Molecular Sciences

Article Title: Histone Deacetylase Inhibition Downregulates Collagen 3A1 in Fibrotic Lung Fibroblasts

doi: 10.3390/ijms141019605

Figure Lengend Snippet: Idiopathic pulmonary fibrosis (IPF) lung tissues ( A ) and primary fibroblasts ( B ) have higher COL3A1 mRNA expression when compared to control samples by real-time RT-PCR. Results are the average of at least three independent experiments (IPF or Control lung tissue samples, n = 3). Bars indicate the mean ± SD. * p < 0.05 compared to the non-IPF control.

Article Snippet: Collagen III antibody (Col3A1 antibody catalog#49-394) was from ProSci Incorporated (Poway, CA, USA); anti-H3Ac (#9671), -H4Ac (#2591), -H4 (#2591) and -β-tubulin (#2128) were from Cell Signaling (Beverly, MA, USA). β-tubulin was used as the loading control for the whole cell lysate; H3 was used as the loading control for nuclear extracts.

Techniques: Expressing, Control, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Histone Deacetylase Inhibition Downregulates Collagen 3A1 in Fibrotic Lung Fibroblasts

doi: 10.3390/ijms141019605

Figure Lengend Snippet: COL3A1 expression in IPF primary fibroblasts with or without suberoylanilide hydroxamic acid (SAHA) treatment. ( A ) Expression of mRNA COL3A1 in IPF cells with or without SAHA by real-time RT-PCR; ( B and C ) Expression of collagen III in IPF whole cell lysate, which indicates intracellular collagen III, by Western blot with 8% SDS-PAGE gel under reducing condition (more information is available in the online supplementary data). Collagen III α1 is at approximately 140 kDa marker . β-tubulin was used as a loading control; C is densitometry by blots like B ; ( D ) Representative pictures of immunofluorescence of IPF cells without (left) and with (right) SAHA, stained with α-SMA (red, bottom) and collagen III (green). Top panels: IPF fibroblasts stained with Col3A1 anti-body (green) indicating intracellular collagen III; nuclei are stained blue (DAPI). Bottom panel: IPF fibroblasts stained with Col3A1 (green) and α-SMA (red); nuclei stained blue (DAPI). The scale bar as shown at the left bottom corner is for 100 μm. The results are the average of at least three independent experiments. The bars indicate the mean ± SD. * p < 0.05 compared to the control.

Article Snippet: Collagen III antibody (Col3A1 antibody catalog#49-394) was from ProSci Incorporated (Poway, CA, USA); anti-H3Ac (#9671), -H4Ac (#2591), -H4 (#2591) and -β-tubulin (#2128) were from Cell Signaling (Beverly, MA, USA). β-tubulin was used as the loading control for the whole cell lysate; H3 was used as the loading control for nuclear extracts.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, SDS Page, Marker, Control, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Histone Deacetylase Inhibition Downregulates Collagen 3A1 in Fibrotic Lung Fibroblasts

doi: 10.3390/ijms141019605

Figure Lengend Snippet: Other histone modification changes and chromatin immunoprecipitation (ChIP) assays demonstrated histone-association changes of COL3A1 with or without SAHA treatment in IPF fibroblasts. ( A ) Western blots of H3K9Ac and H3K27Me3 in IPF primary fibroblasts treated with or without SAHA for 48 h. Total H3 was used as a loading control. Bottom: densitometric analysis of blots, like A ; ( B ) Association changes of histone modifications, H3K9Ac and H3K27Me3, with COL3A1 at the promoter region by ChIP assays. The quantitative ChIP assays were performed to examine the association changes of histone modifications, H3K9Ac (white bars) and H3K27Me3 (black bars), with the promoter region of COL3A1. The bars represent the relative levels of the PCR product of the region of COL3A1 associated with these specific histone modifications under the indicated conditions after immunoprecipitation with these specific antibodies, normalized to input DNA. The bars represent the mean ± SD from the average of at least three independent experiments. * p < 0.05 vs . the untreated control to its own group.

Article Snippet: Collagen III antibody (Col3A1 antibody catalog#49-394) was from ProSci Incorporated (Poway, CA, USA); anti-H3Ac (#9671), -H4Ac (#2591), -H4 (#2591) and -β-tubulin (#2128) were from Cell Signaling (Beverly, MA, USA). β-tubulin was used as the loading control for the whole cell lysate; H3 was used as the loading control for nuclear extracts.

Techniques: Modification, Chromatin Immunoprecipitation, Western Blot, Control, Immunoprecipitation

Journal: International Journal of Molecular Sciences

Article Title: Histone Deacetylase Inhibition Downregulates Collagen 3A1 in Fibrotic Lung Fibroblasts

doi: 10.3390/ijms141019605

Figure Lengend Snippet: Collagen III in the murine model of bleomycin-induced pulmonary fibrosis with or without SAHA treatment. ( A ) Immunohistochemistry with antibody against collagen III in mice lung with saline, bleomycin and bleomycin with SAHA. Representative collagen III stained 5-μm sections of formalin-fixed, paraffin-embedded lung of six- to eight-week-old C57BL mice 28 days after intra-tracheal injection of normal saline (top), bleomycin (Bleo, middle) or bleomycin with SAHA (Bleo + SAHA, bottom) treatment every other day from day 10 to day 28 after bleomycin injury. Immunostaining of collagen III (red) is seen in large areas in the bleomycin only group. Comparatively, there is much less immunostaining of collagen III in the Bleo/SAHA treated group (bottom) compared to the bleomycin group, while little staining is observed in the saline group (a higher magnification of the figures is in the online supplementary data); ( B ) Quantitation of collagen III stained cells in randomly chosen fields relative to the total cell count in the field. The bars represent the mean ± SD of the average of at least three randomly chosen fields. * p < 0.05 vs. the bleomycin group.

Article Snippet: Collagen III antibody (Col3A1 antibody catalog#49-394) was from ProSci Incorporated (Poway, CA, USA); anti-H3Ac (#9671), -H4Ac (#2591), -H4 (#2591) and -β-tubulin (#2128) were from Cell Signaling (Beverly, MA, USA). β-tubulin was used as the loading control for the whole cell lysate; H3 was used as the loading control for nuclear extracts.

Techniques: Immunohistochemistry, Saline, Staining, Formalin-fixed Paraffin-Embedded, Injection, Immunostaining, Quantitation Assay, Cell Counting

Journal: PLOS One

Article Title: A bioactive hydrogel integrating bFGF/VEGFA gene-loaded nanoparticles and platelet-rich plasma for accelerated full-thickness skin wound healing

doi: 10.1371/journal.pone.0350087

Figure Lengend Snippet: (A) Representative Hematoxylin & eosin (H&E) staining images of wound sections; The red arrow refers to the skin appendage. (B) Representative Masson’s trichrome staining images of wound sections; blue color indicates collagen. (C) Representative immunohistochemical images of wound sections showing type-I collagen (COLI) expression. (D) Representative immunohistochemical images of wound sections showing type-III collagen (COLIII) expression. (E) Quantification of total collagen deposition (Masson-stained area %). (F) Quantification of COLI deposition (% positively stained area). (G) Quantification of COLIII deposition (% positively stained area). In E–G, data are mean ± SD from three independent animals (n = 3 mice per group). Statistical significance was evaluated by one-way ANOVA and LSD comparison test. Ns, no significant difference (P > 0.05); *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Anti-bFGF antibody (bsm-34016R, Bioss, China, dilution 1:250), anti-VEGFA antibody (ab46154, Abcam, UK, dilution 1:250), anti-CD 31 antibody (28083–1-AP, Proteintech, USA, dilution 1:250), anti-Ki 67 antibody (ab209897, Abcam, UK, dilution 1:250), anti-Bcl 2 antibody (26593–1-AP, Proteintech, USA, dilution 1:250), anti-Collagen Type I antibody(HY- P81227 , MCE, USA, dilution 1:250), anti- Collagen Type III antibodies (HY- P81101 , MCE, USA, dilution 1:250) were added to the slices and incubated overnight.

Techniques: Staining, Immunohistochemical staining, Expressing, Comparison