Journal: Journal of Asthma and Allergy

Article Title: Integrative Analysis Reveals a miRNA-mRNA Regulatory Network and Potential Causative Agents in the Asthmatic Airway Epithelium

doi: 10.2147/JAA.S331090

Figure Lengend Snippet: Effects of miR-1246 on POSTN and remodeling. ( A ) Western blot and qRT-PCR assays were performed to determine the full time-response curve of periostin. The maximum response was observed with 100 ng/mL IL-13 at 12 h. ( C – E ) The expression of POSTN in BEAS-2B cells transfected with the miR-1246 mimic, miR-1246 inhibitors, or miR-control was measured by Western blot, RT-PCR and immunofluorescence analysis. ( F – M ) RT-PCR was used to examine the effects of miR-1246 on airway remodeling and EMT using miR-1246 mimics ( F – I ) and inhibitors ( J – M ). Similar results were obtained in three independent experiments. *P < 0.05; **P < 0.01.

Article Snippet: The primary anti-periostin antibody (1:200, ab152099, Abcam, USA), anti-fibronectin antibody (1:200, JF0582, Huabio, China), and anti-COL1A1 antibody (1:200, BA0325, Boster, China) were used to incubate the cell climbing slices overnight at 4°C.

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Transfection, Control, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence

Journal: Biochemical and biophysical research communications

Article Title: Overexpression of Snail in retinal pigment epithelial triggered epithelial-mesenchymal transition.

doi: 10.1016/j.bbrc.2014.02.119

Figure Lengend Snippet: Fig. 3. Overexpression of Snail in ARPE-19 cells induced EMT. ARPE-19 cells were transfected with pReceiver-Snail or pReceiver-control for 48 h. QRT-PCR and Immunoblotting were used to examine the expression of Snail, E-cadherin, ZO-1, a-SMA and fibronectin. (A) QRT-PCR analysis showed the increased Snail, fibronectin and a- SMA mRNA expression and decreased E-cadherin and ZO-1 mRNA expression. ⁄⁄P < 0.01 vs pReceiver-control. (B) Immunoblotting confirmed the expression of these EMT markers at protein levels.

Article Snippet: The primary antibodies used were as follows: 1:500 E-cadherin antibody and 1:1000 fibronectin antibody (R&D systems, Inc., USA), 1:1000 Snail antibody (Abcam Ltd., Cambridge, USA), 1:1000 a-SMA antibody (Sigma–Aldrich, MO, USA), 1:1000 ZO-1 antibody (Invitrogen, Carlsbad, CA), and 1:5000 GAPDH (Good HERE, Hangzhou, China).

Techniques: Over Expression, Transfection, Control, Quantitative RT-PCR, Western Blot, Expressing

Journal: Stem Cells

Article Title: Cadherin-11 Influences Differentiation in Human Mesenchymal Stem Cells by Regulating the Extracellular Matrix Via the TGFβ1 Pathway

doi: 10.1093/stmcls/sxac026

Figure Lengend Snippet: Cadherin-11 knockdown reduces the expression of fibronectin. HMSCs were seeded at 1 × 10 4 cells/cm 2 and evaluated after days 1 and 14. ( A ) Western blot analysis of fibronectin on day 1 shows increased expression in cadherin-11-knockdown cells (sh-CDH11) compared with the wild type and scrambled (sh-SCR) controls. GAPDH is shown as a loading control. ( B ) Immunofluorescence micrographs of fibronectin (white) at day 1 also show increased expression in the sh-CDH11 cells compared with the wild type. ( C ) Western blot analysis on day 14 shows that the fibronectin expression persists in sh-CDH11 cells compared with the wild type and sh-SCR. ( D ) Immunofluorescence micrographs of fibronectin (white) at day 14 confirm the increased expression compared to the wild type. Nuclei were counterstained with DAPI (blue). Data are representative of at least 3 independent experiments with similar results. Scale bars represent 100 μm.

Article Snippet: Primary antibodies were against: type VI collagen (rabbit clone, 1:1000; Genetex, GTX109963), fibronectin (rabbit clone, 1:1000; Novus Biologicals, NBP1-91258), cadherin-11 (rabbit clone, 1:1000; Thermo Fisher Scientific, 71-7600), or GAPDH (mouse clone, 1:1000; Santa Cruz Biotechnology, SC-365062).

Techniques: Knockdown, Expressing, Western Blot, Control, Immunofluorescence