Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Louqin Zhisou Decoction Inhibits Mucus Hypersecretion for Acute Exacerbation of Chronic Obstructive Pulmonary Disease Rats by Suppressing EGFR-PI3K-AKT Signaling Pathway and Restoring Th17/Treg Balance

doi: 10.1155/2019/6471815

Figure Lengend Snippet: Effect of LQZS on MUC5AC synthesis and expression in lung tissues of rats with AECOPD. (a) Immunohistochemical staining of MUC5AC in bronchial epithelium (magnification ×100 and ×400). (b) Estimate of MUC5AC expressions through Western blot. β -actin was used as an internal control. The values were shown as proportions of MUC5AC to β -actin optical density. (c) Changes in relative mRNA levels of MUC5AC. The values were expressed as mean ± SD. One-way ANOVA was adopted for statistical analysis. ∗∗ Compared to Control group P < 0.01; ## compared to AECOPD group P < 0.01; && compared to CAM group P < 0.01.

Article Snippet: Then the filters were incubated with primary antibodies, mice monoclonal IgG anti-rat MUC5AC antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat ROR γ t antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat Foxp3 antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat NE antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat MCP-1 antibody (proteintech, China, dilution 1:1000), and rabbit polyclonal IgG anti-rat β -actin (proteintech, China, dilution 1:2000) served as internal control at 4°C overnight.

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Louqin Zhisou Decoction Inhibits Mucus Hypersecretion for Acute Exacerbation of Chronic Obstructive Pulmonary Disease Rats by Suppressing EGFR-PI3K-AKT Signaling Pathway and Restoring Th17/Treg Balance

doi: 10.1155/2019/6471815

Figure Lengend Snippet: Effect of LQZS on repairing Th17/Treg imbalance in lung tissues of rats with AECOPD. Immunohistochemical staining of ROR γ t (a) and Foxp3 (b) proteins in lung tissues (magnification ×100 and ×400). Estimate of ROR γ t (c) and Foxp3 (d) expressions through Western blot. β -actin was used as an internal control. The values were shown as proportions of target proteins to β -actin optical density. Changes in relative mRNA levels of ROR γ t (e), Foxp3 (f), IL-17(g), IL-10 (h) in lung tissues. The values were expressed as mean ± SD. One-way ANOVA was adopted for statistical analysis. ∗ Compared to Control group P < 0.05, ∗∗ P < 0.01; # compared to AECOPD group P < 0.05, ## P < 0.01; & compared to CAM group P < 0.05, && P < 0.01.

Article Snippet: Then the filters were incubated with primary antibodies, mice monoclonal IgG anti-rat MUC5AC antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat ROR γ t antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat Foxp3 antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat NE antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat MCP-1 antibody (proteintech, China, dilution 1:1000), and rabbit polyclonal IgG anti-rat β -actin (proteintech, China, dilution 1:2000) served as internal control at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Western Blot, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Louqin Zhisou Decoction Inhibits Mucus Hypersecretion for Acute Exacerbation of Chronic Obstructive Pulmonary Disease Rats by Suppressing EGFR-PI3K-AKT Signaling Pathway and Restoring Th17/Treg Balance

doi: 10.1155/2019/6471815

Figure Lengend Snippet: Effect of LQZS on NE and MCP-1 synthesis and expressions in lung tissues of rats with AECOPD. Immunohistochemical staining of NE (a) and MCP-1 (b) proteins in lung tissues (magnification ×100 and ×400). Estimate of NE (c) and MCP-1 (d) expressions through Western blot. β -actin was used as an internal control. The values were shown as proportions of target proteins to β -actin optical density. Changes in relative mRNA levels of NE (e) and MCP-1 (f) in lung tissues. The values were expressed as mean ± SD. One-way ANOVA was adopted for statistical analysis. ∗ Compared to Control group P < 0.05, ∗∗ P < 0.01; # compared to AECOPD group P < 0.05, ## P < 0.01; && compared to CAM group P < 0.01.

Article Snippet: Then the filters were incubated with primary antibodies, mice monoclonal IgG anti-rat MUC5AC antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat ROR γ t antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat Foxp3 antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat NE antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat MCP-1 antibody (proteintech, China, dilution 1:1000), and rabbit polyclonal IgG anti-rat β -actin (proteintech, China, dilution 1:2000) served as internal control at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Western Blot, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Trigonella foenum-graecum L. and Psoralea corylifolia L. Improve Erectile Dysfunction in Streptozotocin-Induced Diabetic Rats through Suppression of Oxidative Stress

doi: 10.1155/2022/4187359

Figure Lengend Snippet: Primer sequences used for real-time PCR.

Article Snippet: Rat anti- α -SMA (19245S), anti-Bax (2772S), anti-eNOS (4231S), anti-rabbit IgG (5151P), anti-rat IgG (5257P), and horseradish peroxidase-linked antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Sequencing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Trigonella foenum-graecum L. and Psoralea corylifolia L. Improve Erectile Dysfunction in Streptozotocin-Induced Diabetic Rats through Suppression of Oxidative Stress

doi: 10.1155/2022/4187359

Figure Lengend Snippet: Effect of HLBW on smooth muscle content in the CC of DMED rats. (a) Masson trichrome staining (red indicates the area of smooth muscle, and blue indicates the area of collagen). (b) The ratio of smooth muscle to collagen in Masson-stained CC. (c) RT-PCR for the mRNA expression of α -SMA. (d) Western blot analysis for α -SMA protein expression. (e) Relative expression levels of α -SMA normalized to GAPDH expression. Note. ∗∗∗ P < 0.001, ∗ P < 0.05 vs. control group; ## P < 0.01, # P < 0.05 vs. model group. Data are expressed as the mean ± SD. α -SMA: α -smooth muscle actin.

Article Snippet: Rat anti- α -SMA (19245S), anti-Bax (2772S), anti-eNOS (4231S), anti-rabbit IgG (5151P), anti-rat IgG (5257P), and horseradish peroxidase-linked antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Control

Journal: Experimental and Therapeutic Medicine

Article Title: KDM6A suppresses hepatocellular carcinoma cell proliferation by negatively regulating the TGF-β/SMAD signaling pathway

doi: 10.3892/etm.2020.9000

Figure Lengend Snippet: KDM6A negatively regulates the TGF-β/SMAD signaling pathway. (A) The effect of KDM6A overexpression on the expression of TGF-β, p-smad2, p-smad4, Ki67 and PCNA in Huh7 and LM3 cells was (A) determined by western blotting and (B) semi-quantified. The effect of KDM6A knockdown on the expression of TGF-β, p-smad2, p-smad4, Ki67 and PCNA in YY-8103 and SNU-398 was (C) determined by western blotting and (D) semi-quantified. ** P<0.01 and *** P<0.001 vs. vector or SCR. KDM6A, lysine demethylase 6A; TGF-β, transforming growth factor-β; p, phosphorylated; PCNA, proliferating cell nuclear antigen; SCR, scrambled; sh, short hairpin RNA; ns, not significant.

Article Snippet: Subsequently, the membranes were incubated at 4˚C overnight with the following primary antibodies: Anti-KDM6A (1:1,000; cat. no. orb333886; Biorbyt Ltd.), anti-transforming growth factor (TGF)-β (1:1,000; cat. no. 3711; Cell Signaling Technology, Inc.), anti-phosphorylated (p)-smad2 (1:1,000; cat. no. 18338; Cell Signaling Technology, Inc.), anti-Smad2 (1:1,000; cat. no. 12570-1-AP; ProteinTech Group, Inc.), anti-p-smad4 (1:1,000; cat. no. 10231-8-AP; ProteinTech Group, Inc.), anti-smad4 (1:1,000; cat. no. 10231-1-AP; ProteinTech Group, Inc.), anti-proliferating cell nuclear antigen (PCNA; 1:1,000; cat. no. 10205-2-AP; ProteinTech Group, Inc.), anti-Ki67 (1:1,000; cat. no. 27309-1-AP; ProteinTech Group, Inc.), anti-GAPDH (1:1,000; cat. no. 10494-1-AP; ProteinTech Group, Inc.) and anti-Flag (1:4,000; cat. no. F7425; Sigma-Aldrich; Merck KGaA).

Techniques: Over Expression, Expressing, Western Blot, Plasmid Preparation, shRNA

Journal: Biomaterials

Article Title: Fibrin polymer on the surface of biomaterial implants drives the foreign body reaction.

doi: 10.1016/j.biomaterials.2021.121087

Figure Lengend Snippet: Fig. 9. Macrophages within the fibrous capsule express α-SMA and CD68. PCTFE sections were implanted in WT (A, B) and FibAEK (C, D) mice for 3, 7, and 14 days. Representative images of the capsule sam ples retrieved from WT (n = 3) and FibAEK

Article Snippet: The mouse anti-α-SMA mAb (catalog number MAB1420-SP) was from R&D systems.

Techniques:

Journal: Biomaterials

Article Title: Fibrin polymer on the surface of biomaterial implants drives the foreign body reaction.

doi: 10.1016/j.biomaterials.2021.121087

Figure Lengend Snippet: Fig. 10. Macrophages within the fibrous capsule express collagen. PCTFE sections were implanted in WT and FibAEK mice for 14 days and the cells accumulated within the capsules were isolated as described in the Materials and Methods. (A) The cells were allowed to adhere to the surface of a FluoroDish, fixed, and incubated with Alexa Fluor 568-conjugated phalloidin, anti-α-SMA, and anti-CD68 antibodies. Representative confocal images of myofibroblasts and macrophages isolated from the capsule retrieved from WT mice are shown. Arrowheads point to podosomes seen in macrophages incubated with phalloidin-Alexa Fluor 568. The scale bar is 10 μm. (B) Quantification of cells expressing α-SMA only (myofibroblasts) or both α-SMA and CD68 (macrophages). Approximately 2000–2500 cells isolated from the capsules retrieved from each WT and FibAEK mice were analyzed with ~400 cells present in each field of view. (C) The cells isolated from the capsule obtained from WT mice were incubated with anti-CD11b mAb M1/70 and anti-collagen I mAb. Myofibroblasts shown in the upper panel are indicated by white arrows. The scale bar is 20 μm. (D) Representative confocal images of collagen-expressing macrophages isolated from the WT and FibAEK mice. The cells were labeled for collagen 1 (green) and actin (red). The scale bar is 10 μm. (E) The frequency distribution of fluorescence intensities for collagen I in macrophages isolated from the capsules retrieved from WT and FibAEK mice was expressed as CTCF arbitrary units (A.U). A total of 300 cells from 5 random fields in the samples prepared from WT and FgAEK

Article Snippet: The mouse anti-α-SMA mAb (catalog number MAB1420-SP) was from R&D systems.

Techniques: Capsules, Isolation, Incubation, Expressing, Labeling, Fluorescence

Journal: bioRxiv

Article Title: Neratinib, a clinical drug against breast cancer, protects against atherosclerosis via ASK1 inhibition

doi: 10.1101/2024.11.05.622073

Figure Lengend Snippet: (A) Flow chart of the animal experiment. Ldlr −/− mice were fed a high-cholesterol diet (HCD) for 14 weeks and were concurrently administered with 20 mg/kg Neratinib by oral gavage. (B) Oil Red O staining of the en face aorta. Scale bar: 1 mm, n=10 for each group. (C) Oil Red O staining of aortic sinus. Scale bar: 200 μm, n=10 for each group. (C and D) H&E and Masson staining of aortic sinus cryosections. Necrotic core size and collagen content in plaques was quantified. Scale bar: 200 μm, n=10 for each group in C and n=9 for vehicle group and n=10 for Ner group in D. (F) Immunofluorescence staining of CD68 and α-SMA in the aortic sinus. CD68 positive infiltrated macrophages and α-SMA positive smooth muscle cell content were quantified. Scale bar: 200 μm, n=10 for each group. All data are presented as the means ± SEMs. Data of B are analyzed using Kolmogorov-Smirnov test. Data of C, E and α-SMA + area in F were analyzed using unpaired Student’s t-test. Data of D and CD68 + area in F were analysis using unpaired Student’s t-test with Welch’s correction.

Article Snippet: The primary antibodies used were rat anti-mouse CD68 antibody (FA-11; Bio-Rad, USA; 1:500 dilution) and rabbit anti-mouse SMA antibody (14395-1-AP; Proteintech, USA; 1:500 dilution).

Techniques: Staining, Immunofluorescence

Journal: bioRxiv

Article Title: Neratinib, a clinical drug against breast cancer, protects against atherosclerosis via ASK1 inhibition

doi: 10.1101/2024.11.05.622073

Figure Lengend Snippet: Eight-week-old female Ldlr −/− mice were fed a high-fat high-cholesterol diet for 14 weeks and were orally administered with vehicle or 20 mg/kg Neratinib. n=7 in each group. (A) Oil Red O staining of the en face aorta. Scale bar: 1 mm. (B) Oil Red O staining of aortic sinus cryosections. Scale bar: 20 μm. (C and D) H&E and Masson staining of aortic sinus sections to indicate the necrotic core area and collagen content. Scale bar:250 μm. (E) Immunofluorescence staining of CD68 and α-SMA in the aortic sinus plaques to indicate infiltrated macrophages and smooth muscle cell content. Scale bar: 200 μm. All data are presented as the means ± SEMs. Data of A-E except α-SMA in E were analyzed with unpaired Student’s t-test. Data of α-SMA in E were analyzed unpaired Student’s t-test with Welch’s correction.

Article Snippet: The primary antibodies used were rat anti-mouse CD68 antibody (FA-11; Bio-Rad, USA; 1:500 dilution) and rabbit anti-mouse SMA antibody (14395-1-AP; Proteintech, USA; 1:500 dilution).

Techniques: Staining, Immunofluorescence

Journal: bioRxiv

Article Title: Neratinib, a clinical drug against breast cancer, protects against atherosclerosis via ASK1 inhibition

doi: 10.1101/2024.11.05.622073

Figure Lengend Snippet: (A) Scheme of study design. (B) Graphic of the benefit of combination therapy (C) Gross image and Oil Red O-stained image of en face aorta. Scale bar: 1 mm, n=10 for Neratinib group, n=17 for Rosuvastatin, and n=19 for vehicle and combination group in gross image and n=10 for Neratinib group, n=18 for Rosuvastatin, and n=20 for vehicle and combination group in en face ORO. (D) Oil Red O staining of aortic sinus. Scale bar: 200 μm, n=10 for Neratinib group, n=18 for Rosuvastatin, and n=20 for vehicle and n=19 for combination group. Nile red staining of neutral lipids in aortic sinus. Scale bar: 50 μm, n=8 for Neratinib group, n=17 for Rosuvastatin, n=20 for vehicle and n=19 for combination group. (E) H&E staining of aortic sinus to indicate the necrotic core area. Scale bar: 625 μm, n=10 for Neratinib group, n=18 for Rosuvastatin, and n=20 for vehicle combination group. (F) Immunofluorescence staining of CD68 and α-SMA in the aortic sinus. Scale bar: 50 μm, n=10 for Neratinib group, n=119 for Rosuvastatin, n=20 for vehicle and combination group. (G and H) Olink proteomics illustrating differentially secreted inflammation associated cytokines in the combination group vs rosuvastatin group, n=8 for Rosuvastatin and n=10 combination group. Abbreviations: ROSU: Rosuvastatin, Ner: Neratinib. All data are presented as the means ± SEMs. Data of C, ORO in D, E and F are analysed using Kruskal-Wallis test. Data of Nile Red staining in D are analyzed using Brown-Forsythe and Welch ANOVA test. Data of H were analysed using unpaired Student’s t-test with Welch’s correction in IL-7, CCL2, CCL4, and CSF2, and unpaired Student’s t-test in CXCL19. Data of IL-16 in H were analyzed using Mann-Whitney test.

Article Snippet: The primary antibodies used were rat anti-mouse CD68 antibody (FA-11; Bio-Rad, USA; 1:500 dilution) and rabbit anti-mouse SMA antibody (14395-1-AP; Proteintech, USA; 1:500 dilution).

Techniques: Staining, Immunofluorescence, MANN-WHITNEY

Journal: Cell Death and Differentiation

Article Title: Caspase-10: a molecular switch from cell-autonomous apoptosis to communal cell death in response to chemotherapeutic drug treatment

doi: 10.1038/cdd.2017.164

Figure Lengend Snippet: Lack of Caspase-10 leads to TNF-α production in response to 5FU and FADD-independent apoptosis. (a) (Left) Caspase-8 activity was measured in response to 5FU in HCT.shC10 cells. (Right) Double knockdown clones, HCT.shC10.shctrl, HCT.shC10.shC8 and HCT.shC10.shFADD were tested for apoptosis in response to 5FU. Data are plotted as mean±S.E.M. (n≥3). (b) 5FU-induced apoptosis was measured in HCT.shC10 cells in the presence of TNF-α and TNFR1-blocking antibodies. IgG1 and IgG2a antibodies were used as isotype controls, respectively. Data are plotted as mean±S.E.M. (n≥3). (c) TNF-α was measured in supernatants of 5FU-treated HCT.shC10 cells. Data are plotted as mean±S.E.M. (n=3). (d) HCT.shC10 cells after 5FU treatment were analysed by western blot for IκB-α and phospho-IκB-α. (e) HCT.shC10 cells expressing IκB-SR (Ad.IκB-SR) were treated with 5FU, after which TNF-α and apoptosis were measured. Ad.EGFP served as control. Data are plotted as mean±S.E.M. (n≥3). (f) HCT116 cells silenced for both caspase-10 and caspase-8 were treated with 5FU and TNF-α was measured. Data are plotted as mean±S.E.M. (n≥5). (g) Western blot for cFLIP in HCT116 and HCT.shC10 cells after 5FU treatment. (h) EGFP, cFLIPS, cFLIPL, cFLIPp43 and cFLIPD376N (FLIPDN) were expressed in HCT116 cells, and then the cells treated with 5FU before TNF-α levels were measured. Data are plotted as mean±S.E.M. (n≥2). (i) HCT.shC10 cells were silenced for cFLIP (Ad.shFLIP). TNF-α in cell culture supernatants was then determined after 5FU stimulation. Data are plotted as mean±S.E.M. (n=2). (j) Either cFLIPL or cFLIPp43 were expressed in HCT.shC8 cells. After 5FU treatment, TNF-α was measured. Data are plotted as mean±S.E.M. (n≥3). (k) Precipitates from a caspase-8 IP were probed for cFLIP, FADD and caspase-8. The lysates were from HCT116 cells overexpressing cFLIPL and treated with 5FU. Cells overexpressing EGFP were used as controls. Input controls are shown on the left. (l) End point tumour volumes of HCT.shC10 xenografts treated with 5FU are depicted in relation to starting volumes (set to 100). (animal numbers/group: n=6/shctrl, n=3/shC10].

Article Snippet: Sheep anti-CuZnSOD (The Binding Site, Birmingham, UK), goat and mouse anti-actin (Santa Cruz Biotechnology), rabbit anti-TRAIL-R2 (Cell Signaling Technology, Danvers, MA, USA), mouse anti-caspase-3 (Imgenex, Minneapolis, MN, USA), mouse anti-caspase-8 (Cell Signaling Technology, Santa Cruz Biotechnology and BD Biosciences, Franklin Lakes, NJ, USA), mouse anti-XIAP (BD Biosciences), mouse anti-cFLIP (AdipoGen, San Diego, CA, USA), goat anti-cIAP1 (R&D Systems), rat anti-cIAP2 (Enzo Life Sciences), mouse anti-caspase-9 (Novus Biologicals, Minneapolis, MN, USA), mouse anti-caspase-2 (BD Biosciences), mouse anti-CD178 (BD Biosciences), mouse anti-TNFR1 (Santa Cruz Biotechnology), rabbit anti-TRAIL (Peprotech), rabbit anti-TRAIL-R1 (Santa Cruz Biotechnology), mouse anti-TNFR1 (Hycult Biotech, Uden, Netherlands), mouse anti-FADD (Millipore, Billerica, MA, USA), rabbit anti-Bid (R&D Systems), mouse anti-RIP1 (BD Biosciences), mouse anti-p53 (BD Biosciences), mouse anti-caspase-10 (MBL, Woburn, MA, USA), rabbit anti-CD95 (Santa Cruz Biotechnology), mouse anti-TRAF2 (Santa Cruz Biotechnology), rabbit anti-TRADD (Cell Signaling Technology), rabbit anti-CRADD/RAIDD (Cell Signaling Technology), rabbit anti-NEMO (Santa Cruz Biotechnology), mouse anti-Myc (Santa Cruz Biotechnology), mouse anti-V5 (Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti-I κ B- α (Cell Signaling Technology), rabbit anti-Phospho-I κ B- α (Cell Signaling Technology), rabbit anti-Phospho-ATR (GeneTex, Irvine, CA, USA), rabbit anti-ATR (Bethyl Laboratories, Montgomery, TX, USA), rabbit anti-Phospho-ATM (Cell Signaling Technology), mouse anti-ATM (Sigma), rabbit anti-Phospho-Chk1 (Cell Signaling Technology), mouse anti-Chk1 (Cell Signaling Technology), mouse anti-TNF- α (Biolegend, San Diego, CA, USA), IgG1 Isotype antibody (BD Biosciences), IgG2a Isotype antibody (Biolegend), IgG2b Isotype antibody (Biolegend), mouse anti-Smac/DIABLO antibody (Abcam, Cambridge, UK), FITC-conjugated F(ab‘)2 anti-mouse IgG2a antibody (Southern Biotechnology, Birmingham, AL, USA), FITC-conjugated F(ab‘)2 anti-mouse IgG2b antibody (Southern Biotechnology), mouse anti-cytochrome c (BD Biosciences).

Techniques: Activity Assay, Knockdown, Clone Assay, Blocking Assay, Western Blot, Expressing, Control, Cell Culture

Journal: Molecular Therapy. Nucleic Acids

Article Title: miR-133b Downregulation Reduces Vulnerable Plaque Formation in Mice with AS through Inhibiting Macrophage Immune Responses

doi: 10.1016/j.omtn.2019.04.024

Figure Lengend Snippet: Downregulated miR-133b Increases the Positive Rate of Macrophages and Decreases the Positive Rate of VSMCs in the AS Mouse Models (A) The immunohistochemical staining of CD68 in each group (scale bar, 25 μM). (B) The immunohistochemical staining of α-SMA in each group (scale bar, 25 μM). AS, atherosclerosis; VSMC, vascular smooth muscle cell; miR-133b, microRNA-133b.

Article Snippet: Then the sections were blocked with 10% normal goat serum for 20 min at room temperature, and they were incubated with primary antibodies rabbit polyclonal antibody to α-SMA (dilution ratio of 1:100–300, A03744, Boster Biological Technology, Wuhan, China) and CD68 (dilution ratio of 1:2,000, ab125212, Abcam, Cambridge, MA, USA) overnight at 4°C, with PBS serving as the NC.

Techniques: Immunohistochemical staining, Staining

Journal: Molecular Therapy. Nucleic Acids

Article Title: miR-133b Downregulation Reduces Vulnerable Plaque Formation in Mice with AS through Inhibiting Macrophage Immune Responses

doi: 10.1016/j.omtn.2019.04.024

Figure Lengend Snippet: Quantitative Analysis for the Positive Rates of CD68 and α-SMA in Carotid Artery of Mice Detected by Immunohistochemistry

Article Snippet: Then the sections were blocked with 10% normal goat serum for 20 min at room temperature, and they were incubated with primary antibodies rabbit polyclonal antibody to α-SMA (dilution ratio of 1:100–300, A03744, Boster Biological Technology, Wuhan, China) and CD68 (dilution ratio of 1:2,000, ab125212, Abcam, Cambridge, MA, USA) overnight at 4°C, with PBS serving as the NC.

Techniques: