Journal: Indian Journal of Pharmaceutical Sciences

Article Title: Protective Effects of Ginsenoside Rb1 in Rats with Diabetic Cardiomyopathy

doi: 10.36468/pharmaceutical-sciences.967

Figure Lengend Snippet: Fig. 4: Ginsenoside Rb1 attenuated inflammation and suppressed the HB-EGF pathway in DCM rats, (A): RNA expressions of IL6 as determined by quantitative real-time RT-PCR; (B): Protein expressions of HB-EGF in serum as determined by ELISA and (C): Protein expressions of HB-EGF in heart tissue as determined by Western blot Note: Compared to NC, ***p<0.001 and compared to DCM, ###p<0.001

Article Snippet: Human Heparin-Binging Epidermal Growth Factor (HB-EGF) Enzyme-Linked Immunosorbent Assay (ELISA): A Human HB-EGF ELISA kit (Elabscience, China) was used to test for the activity of HB-EGF in plasma, according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: International Journal of General Medicine

Article Title: Characterization of Gastric Mucinous Carcinoma: Clinicopathological Insights and Vascular Features via MSCT

doi: 10.2147/IJGM.S567353

Figure Lengend Snippet: Immunohistochemistry (IHC) detection of vascular endothelial growth factor A (VEGF-A), hypoxia-inducible factor-1α (HIF-1α), and epidermal growth factor (EGF) in carcinomas and corresponding paracancerous normal tissues of MGC, LEMPC, and NMGC. ( A – C ) Representative immunohistochemical staining of VEGF-A ( A ), HIF-1α ( B ), EGF( C ) in carcinomas and normal tissues (×400, bar = 50μm). ( D – F ) IHC score differences (cancer tissue score - normal tissue score) for VEGF-A ( D ) HIF-1α ( E ) EGF ( F ) among the three groups. Statistical significance is indicated as follows: P < 0.05; P < 0.001.

Article Snippet: Sections were stained for CD34 (1:2500, Abcam, ab81289), VEGF-A (1:800, Proteintech, 66828-1-IG), HIF-1α (1:200, Proteintech, 20960-1-AP), and EGF (1:200, Proteintech, 27141-1-AP).

Techniques: Immunohistochemistry, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Oxidized Mitochondrial DNA Engages TLR9 to Activate the NLRP3 Inflammasome in Myelodysplastic Syndromes

doi: 10.3390/ijms24043896

Figure Lengend Snippet: Ox-mtDNA activates pyroptosis. ( A ) Log10-transformed, glucose-adjusted ox-mtDNA levels from Low Risk (LR) MDS PB (n = 100) and BM plasma (n = 70), and Normal PB (n = 30). ( B ) Western blot of U937 and SKM1 cells treated with 10-fold increasing isolated mtDNA of the ND1 gene region amplified with oxidized guanosine (ox-mtDNA) for 2 h to induce Caspase-1 (arrow depicts cleaved fragment) and phosphorylated NFκB to establish dosage (representative blot of n = 3). Remaining figures, ox-mtDNA treatment is 50 ng/mL ox-mtDNA for 2 h unless otherwise stated. ( C ) Western blot of SKM1 and U937 cells treated with ox-mtDNA showing, cleavage of caspase-1 (arrow depicts cleaved fragment), and IL-1β (arrow depicts cleaved fragment) demonstrating inflammasome activation (representative blot of n = 3). ( D ) Fold change of caspase-1 activity quantified by Caspase-1 Glo ® assay in SKM1 and U937 cells treated with ox-mtDNA, (mean ± SEM of n = 5). ( E ) Fold change of LDH media release (measurement of cell death), quantified by LDH-Glo™ Cytotoxicity Assay, of SKM1 and U937 cells treated with ox-mtDNA (mean ± SEM of n = 5). ( F ) Fold change of caspase-1 activity quantified by Caspase-1 Glo ® assay in SKM1 cells pretreated with either CRISPR KO (pooled guides) for NLRP3 or 10 uM MCC950 for 48 h prior to treatment with ox-mtDNA (mean ± SEM of n = 4). ( G ) Representative confocal IF micrographs showing increased ASC specks in ox-mtDNA stimulated cells compared to untreated controls [DAPI (blue), ASC (green) (×2520)]. ( H ) Quantification of 1-to-2 μm ASC speck IF, at least 200 cells counted per group, (mean ± SEM of n = 3). ( I ) Immunoblot of ASC following chemical crosslinking, cells treated with ox-mtDNA or positive control LAN (LPS + ATP + Nigericin), arrow indicates oligomers. ( J ) Western Blot for PARP and Caspase-3. Western blot of SKM1 and U937 cells treated with ox-mtDNA, and an apoptosis positive control (A431 EGF Stimulated) showing induction of PARP1, cleavage of caspase-3 (arrow depicts cleaved fragments, representative blots of n = 3). ( K ) Fold change of caspase-1 activity quantified by Caspase-1 Glo ® assay and LDH media release quantified by LDH-Glo™ Cytotoxicity Assay in primary Normal BM-MNC cells treated with ox-mtDNA (mean ± SEM of n = 3). ( L ) Colony formation assay for hematopoiesis in Healthy BM-MNCs treated with 50 ng/mL ox-mtDNA for 14 days (representative picture). ( M ) Quantification of various hematopoietic progenitor colonies in response to treatment with 50 ng/mL of ox-mtDNA for 14 days (mean ± SEM of n = 4). In this figure, significance was assessed by either paired t -test (comparison between two groups only) or ordinary one-way ANOVA with multiple comparison analysis in GraphPad Prism. p values are shown as asterisk: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Apoptosis positive control was A431 Whole Cell Lysate EGF Stimulated (Rockland Immunochemicals, Inc., Limerick, PA, USA).

Techniques: Transformation Assay, Clinical Proteomics, Western Blot, Isolation, Amplification, Activation Assay, Activity Assay, Glo Assay, Cytotoxicity Assay, CRISPR, Positive Control, Colony Assay, Comparison

Journal: Molecular Medicine Reports

Article Title: Characterization of bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice: Evaluation of the feasibility of dimethyloxallyl glycine as a mobilization agent

doi: 10.3892/mmr.2016.4945

Figure Lengend Snippet: Paracrine capacity of NBM-MSCs and DBM-MSCs. (A) Quantification of TGF concentration in the culture supernatant of NBM-MSCs and DBM-MSCs. There was no significant difference between the expression levels (P>0.05). (B) Quantification of PDGF concentration in the culture supernatant of NBM-MSCs and DBM-MSCs. No significant difference was identified between them (P>0.05). TGF and PDGF concentrations were detected using enzyme-linked immunosorbent assays. Data are presented as the mean ± standard deviation. BM-MSCs, bone marrow-derived mesenchymal stem cells from normal saline-treated mice; DBM-MSCs, bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice; TGF, transforming growth factor; PDGF, platelet-derived growth factor.

Article Snippet: Mouse TGF (β IG-H3) and PDGF ELISA kits (Wuhan Boster Biological Co., Ltd., Wuhan, China) were used, according to the manufacturer's protocol.

Techniques: Concentration Assay, Expressing, Standard Deviation, Derivative Assay, Saline

Journal: Cells

Article Title: Effect of Expansion Media on Functional Characteristics of Bone Marrow-Derived Mesenchymal Stromal Cells

doi: 10.3390/cells12162105

Figure Lengend Snippet: Surface antigen expression by MSCs grown in media 1, 4, 7, 10, and 13. MSCs grown in media 1 (100% αMEM+8%PL; black), 4 (95% αMEM+8%PL + 5% StemMACS TM ; blue), 7 (50% αMEM+8%PL + 50% StemMACS TM ; green), 10 (5% αMEM+8%PL + 95% StemMACS TM ; red), and 13 (100% StemMACS TM ; violet) were analyzed for the expression of different surface antigens. These included metabolism-related markers GLUT1, GLUT3, GLUT4, FGFR1, FGFR2, FGFR3, PDGFRA, PDGFRB, INSR, IGF1R, IGF2R, STRA6, NGFR, and EGFR ( A ), cell adhesion-related markers CD29, CD49a, CD49c, CD49d, CD49e, CD49f, CD51, CD61, CD31, CD44, and CD146 ( B ), tetraspanins CD9, CD63, and CD81 ( C ), as well as the additional markers CD3, CD36, CD362, CD10, CD13, MHC I, and MSCA1 ( D ). Data are presented as mean ± SD and N ≥ 3 independent experiments were performed. Statistically significant differences are depicted as follows: *: p < 0.05; **: p < 0.01; ***: p < 0.001.

Article Snippet: The following antibodies were used for the analyses: CD3 (clone SK7; BD Biosciences, Franklin Lakes, NJ, USA or BioLegend, San Diego, CA, USA), CD9 (clone M-L13; BD Biosciences), CD10 (clone HI10a; BD Biosciences), CD13 (clone WM15; BD Biosciences), CD14 (clone MØP9; BD Biosciences or clone M5E2 or HCD14; both from BioLegend), CD29 (clone TS2/16; Thermo Fisher Scientific Inc., Waltham, MA, USA), CD31 or platelet/endothelial cell adhesion molecule 1 (PECAM-1) (clone WM59; BD Biosciences or BioLegend), CD34 (clone 8G12; BD Biosciences), CD36 (clone AC106; Miltenyi Biotec B.V. & Co. KG), CD44 or homing cell adhesion molecule (HCAM) (clone G44-26; BD Biosciences), CD45 (clone 2D1 or HI30; both from BD Biosciences), CD49a (clone SR84; BD Biosciences), CD49c (clone C3 II.1; BD Biosciences), CD49d (clone 9F10; BD Biosciences), CD49e (clone IIA1; BD Biosciences), CD49f (clone GoH3; BD Biosciences), CD51 (clone NKI-M9; BioLegend), CD61 (clone VI-PL2; BD Biosciences), CD63 (clone H5C6; BD Biosciences), CD73 (clone AD2; BD Biosciences), CD81 (clone JS-81; BD Biosciences), CD90 (clone 5E10; BD Biosciences), CD105 (clone SN6; Bio-Rad AbD Serotec GmbH, Puchheim, Germany or clone 266; BD Biosciences), CD140a or platelet-derived growth factor receptor (PDGFR)A (clone 16A1; BioLegend), CD140b or PDGFRB (clone 28D4; BD Biosciences), CD146 or melanoma cell adhesion molecule (MCAM) (clone P1H12; BD Biosciences), CD220 or insulin receptor (INSR) (clone 3B6/IR; BD Biosciences), CD221 or IGF-I receptor (IGF1R) (clone 1H7; BioLegend), CD222 or IGF-II receptor (IGF2R) (clone QA19A18; BioLegend), CD271 or nerve growth factor receptor (NGFR) (clone ME20.4; BioLegend), CD331 or fibroblast growth factor receptor (FGFR)1 (clone M17A3; Novus Biologicals, LLC, Centennial, CO, USA), CD332 or FGFR2 (clone #98725; R&D Systems, Inc., Minneapolis, MN, USA), CD333 or FGFR3 (clone #136334; R&D Systems, Inc.), CD362 or Syndecan-2 (clone #305515R; R&D Systems, Inc.), epidermal growth factor receptor (EGFR) (clone AY13; BioLegend), glucose transporter (GLUT)1 (clone 202915; BD Biosciences), GLUT3 (clone #202017; R&D Systems, Inc.), GLUT4 (clone #925932; R&D Systems, Inc.), MHC I (clone G46-2.6; BD Biosciences), MHC II (clone Tu39; BD Biosciences), mesenchymal stromal cell antigen-1 (MSCA1) (clone W8B2; Miltenyi Biotec B.V. & Co. KG) and stimulated by retinoic acid 6 (STRA6) (clone #496613; R&D Systems, Inc.).

Techniques: Expressing