Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: FEBS letters

Article Title: miR-190a inhibits epithelial-mesenchymal transition of hepatoma cells via targeting the long non-coding RNA treRNA.

doi: 10.1016/j.febslet.2015.11.024

Figure Lengend Snippet: Fig. 3. Suppression of treRNA by miR-190a inhibits EMT in HCC. (A) 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or sitreRNA2), 20 nM miR-190a, or NC RNA was transfected into Huh-7 or HepG2 cells. EMT related genes were quantified 72 h post-transfection as indicated. (B) Cells were treated with antagomiR (anti-miR- 190a) or NC RNA as in (A). The mRNA level of different EMT genes was quantified and normalized to b-actin. (C) Cell lysates were blotting with EMT marker E-cadherin (E-cad), Claudin-1 (Claud), Vimentin (Vimen), N-cadherin (N-cad), SNAI1 (Snail). b-Actin was used as endogenous control. The EMT marker expression level in was calculated using Image J and presented as histogram (right panel). (D) Cells were treated with TGF-b for 12 h and transfected with 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or si-treRNA2) or NC RNA subsequently. The morphological change was captured 24 h after transfection.

Article Snippet: Membranes were blocked by 5% BSA or 5% non-fat milk for 3 h at 25 C and incubated with primary antibody against Ago2 (#2897, Cell Signaling, US), Dicer (#5362, Cell Signaling), E-Cadherin (#3195, Cell Signaling), Claudin-1 (#13255, Cell Signaling), Vimentin (#5741, Cell Signaling), N-Cadherin (#13116, Cell Signaling), SNAI1 (#3897, Cell Signaling), or b-actin (C-4, Santa cruz, US) for 12 h at 4 C. Membranes were incubated with secondary antibody diluted in Tris-buffered saline supplemented with 0.5% Tween-20 and 5% BSA or 5% non-fat milk for 1 h at 25 C. The signal was detected with the enhanced chemiluminescence system (Pierce, US) and exposed to X-ray film (Kodak, US).

Techniques: Transfection, Marker, Control, Expressing