Journal: International Journal of Molecular Medicine

Article Title: BMS-345541 inhibits airway inflammation and epithelial-mesenchymal transition in airway remodeling of asthmatic mice

doi: 10.3892/ijmm.2018.3762

Figure Lengend Snippet: BMS-345541 inhibits epithelial-mesenchymal transition in asthma. mRNA levels of E-cadherin and vimentin in mouse lung tissues were measured by (A) reverse transcription-quantitative polymerase chain reaction analysis. (B) Protein levels were measured by western blot analysis for (C) E-cadherin and (D) vimentin. Data are presented as the mean ± standard deviation (n=8 per group). # P<0.01 and ## P<0.001, OVA group compared with the control group; * P<0.05 and ** P<0.01, OVA + BMS-345541 group compared with the OVA group. OVA, ovalbumin; DMSO, dimethyl sulfoxide.

Article Snippet: The reagents used were as follows: OVA (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), acetylcholine chloride (Ach; Sigma-Aldrich; Merck KGaA), phosphate-buffered saline (PBS; Sigma-Aldrich; Merck KGaA), sodium pentobarbital (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China), BMS-345541 (Abcam, Cambridge, MA, USA), E-cadherin antibody (cat. no. ab76055; Abcam), vimentin antibody (cat. no. ab92547, Abcam), β-actin antibody (cat. no. ab179467; Abcam), glyceraldehyde 3-phosphate dehydrogenase (GADPH; Sigma-Aldrich; Merck KGaA), dimethyl sulfoxide (DMSO; Sigma-Aldrich; Merck KGaA), diaminobenzidine (DAB; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), an enzyme-linked immunosorbent assay (ELISA) kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA), Pierce Enhanced Chemiluminescence (ECL) western blot substrate (Thermo Fisher Scientific, Inc., Waltham, MA, USA), a reverse transcription kit (Promega Corporation, Madison, WI, USA), a quantitative polymerase chain reaction (qPCR) kit (TransStart Green; Beijing Transgen Biotech Co., Ltd., Beijing, China), a bicinchoninic acid (BCA) protein assay kit and radioimmunoprecipitation assay (RIPA) lysis buffer (Beijing ComWin Biotech Co., Ltd., Beijing, China).

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation, Control

Journal: World Journal of Stem Cells

Article Title: Long non-coding RNA SNHG16 promotes human placenta-derived mesenchymal stem cell proliferation capacity through the PI3K/AKT pathway under hypoxia

doi: 10.4252/wjsc.v14.i9.714

Figure Lengend Snippet: SNHG16 was a potential promotor of human placenta-derived mesenchymal stem cell proliferation ability. A: Cell cycle analysis of human placenta-derived mesenchymal stem cells (hP-MSCs) under hypoxic culture via flow cytometry; B: Western blot analysis of AKT phosphorylation in hP-MSCs exposed to hypoxia; C: Circos plot of the long non-coding RNAs (lncRNAs)-messenger (m)RNA co-expression network. The outermost circle is the autosomal distribution. The second and third circles are the distribution of differentially expressed lncRNAs on chromosomes. The red line represents upregulation, and the green line represents downregulation. Higher bars indicate a greater number of differential genes in the interval. The fourth and fifth circles are the distribution of differentially expressed genes on chromosomes, with the same interpretation as lncRNA; D: Part of lncRNA-mRNA interaction network analysis visualized using the Cytoscape software; E: Part of the association analysis of differentially expressed lncRNAs and transcription factors; F: Effects of hypoxia on the expression of SNHG16 in hP-MSCs by quantitative reverse transcription polymerase chain reaction. Data are presented as means ± standard deviation. b P < 0.01.

Article Snippet: Total RNA of cells was obtained using Trizol reagent (Invitrogen, Carlsbad, CA, United States) following the manufacturer’s protocol; the concentration of total RNA was quantified using a NanoDrop-2000 (Thermo Fisher Scientific, Waltham, MA, United States). cDNA was synthesized by reverse transcription reaction using a commercial lncRNA quantitative reverse transcription polymerase chain reaction (PCR) Starter Kit (RiboBio, Guangzhou, China).

Techniques: Derivative Assay, Cell Cycle Assay, Flow Cytometry, Western Blot, Phospho-proteomics, Expressing, Software, Reverse Transcription, Polymerase Chain Reaction, Standard Deviation

Journal: World Journal of Stem Cells

Article Title: Long non-coding RNA SNHG16 promotes human placenta-derived mesenchymal stem cell proliferation capacity through the PI3K/AKT pathway under hypoxia

doi: 10.4252/wjsc.v14.i9.714

Figure Lengend Snippet: Knockdown of SNHG16 attenuated the proliferation ability of human placenta-derived mesenchymal stem cells. A: Quantitative reverse transcription polymerase chain reaction analysis of relative SNHG16 expression after transfection of SNHG16 short hairpin RNA (sh- SNHG16 ) and the corresponding controls (sh-NC) in human placenta-derived mesenchymal stem cells; B: Cell proliferation capacity evaluated by cell counting kit-8 assay; C: Cell cycle measured by flow cytometry; D: The G1 to S phase transition-related proteins and p-AKT detected by western blot analysis. Data are presented as means ± standard deviation. b P < 0.01.

Article Snippet: Total RNA of cells was obtained using Trizol reagent (Invitrogen, Carlsbad, CA, United States) following the manufacturer’s protocol; the concentration of total RNA was quantified using a NanoDrop-2000 (Thermo Fisher Scientific, Waltham, MA, United States). cDNA was synthesized by reverse transcription reaction using a commercial lncRNA quantitative reverse transcription polymerase chain reaction (PCR) Starter Kit (RiboBio, Guangzhou, China).

Techniques: Knockdown, Derivative Assay, Reverse Transcription, Polymerase Chain Reaction, Expressing, Transfection, shRNA, Cell Counting, Flow Cytometry, Sublimation, Western Blot, Standard Deviation

Journal: World Journal of Stem Cells

Article Title: Long non-coding RNA SNHG16 promotes human placenta-derived mesenchymal stem cell proliferation capacity through the PI3K/AKT pathway under hypoxia

doi: 10.4252/wjsc.v14.i9.714

Figure Lengend Snippet: SNHG16 overexpression resulted in activation of the PI3K/AKT pathway and a significant enhancement in the proliferative rate of human placenta-derived mesenchymal stem cells. A: Quantitative reverse transcription polymerase chain reaction analysis of relative SNHG16 expression after transfection of lentivirus overexpressing SNHG16 ( SNHG16 -OE) and the corresponding empty vector in human placenta-derived mesenchymal stem cells; B: Cell proliferation after SNHG16 overexpression was evaluated by cell counting kit-8 assay; C: Cell cycle distribution after SNHG16 overexpression was evaluated by flow cytometry; D: The expression levels of CDK2, CDK4, CDK6, cyclin E1, cyclin D1, and phosphorylated AKT. Data are presented as the means ± SD obtained from three separate experiments. b P < 0.01.

Article Snippet: Total RNA of cells was obtained using Trizol reagent (Invitrogen, Carlsbad, CA, United States) following the manufacturer’s protocol; the concentration of total RNA was quantified using a NanoDrop-2000 (Thermo Fisher Scientific, Waltham, MA, United States). cDNA was synthesized by reverse transcription reaction using a commercial lncRNA quantitative reverse transcription polymerase chain reaction (PCR) Starter Kit (RiboBio, Guangzhou, China).

Techniques: Over Expression, Activation Assay, Derivative Assay, Reverse Transcription, Polymerase Chain Reaction, Expressing, Transfection, Plasmid Preparation, Cell Counting, Flow Cytometry