Journal: PLoS ONE

Article Title: Popeye Domain Containing 1 (Popdc1/Bves) Is a Caveolae-Associated Protein Involved in Ischemia Tolerance

doi: 10.1371/journal.pone.0071100

Figure Lengend Snippet: (a) Western blot analysis of gradient fractions. Fraction 1 and fraction 12 represent the lowest and highest density, respectively. [+/+], WT; [−/−], Popdc1-null; MβCD, methyl-β-cyclodextrin; I/R, ischemia/reperfusion; Cx43, connexin43. The molecular weights of the reactive bands were ∼68 kDa, ∼22 kDa, and ∼43 kDa, for Popdc1, Cav3, and Cx43, respectively. (b) The relative distribution of cholesterol and Cav3 throughout gradient fractions 1–9. *P<0.05, Cav3 in fraction 5, Popdc1-null vs. WT; # P<0.05, cholesterol in fraction 5, Popdc1-null vs. WT.

Article Snippet: For confocal immune-histochemistry, paraformaldehyde-fixed cryosections were incubated with anti-Popdc1 (sc-49889), anti-Cav3 (sc-5310), anti-vinculin (sc-5573) and anti-connexin43 (sc-9059) primary antibodies (purchased from Santa Cruz Biotechnology) and Cy3-conjugated or DyLight 488-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Premature Osteoblast Clustering by Enamel Matrix Proteins Induces Osteoblast Differentiation through Up-Regulation of Connexin 43 and N-Cadherin

doi: 10.1371/journal.pone.0023375

Figure Lengend Snippet: At time point 5 days post seeding, primary human osteoblasts were stained for connexin 43 or N-cadherin (red), and nuclei (blue). Expression of connexin 43 and N-cadherin significantly increased on cell membranes of EMD coated samples. (bar = 50 µm).

Article Snippet: Cells were permealized with Triton X-100 for 5 minutes, followed by staining with antibodies against connexin 43 (sc-9059) or N-cadherin (sc-7939) (Santa Cruz Biotechnologies Inc., Heidelberg, Germany) antibodies, followed by a goat anti-rabbit IgG conjugated to texas red at a dilution of 1∶200 in 0.5% PBS/bovine serum albumin (Invitrogen).

Techniques: Staining, Expressing

Journal: PLoS ONE

Article Title: Premature Osteoblast Clustering by Enamel Matrix Proteins Induces Osteoblast Differentiation through Up-Regulation of Connexin 43 and N-Cadherin

doi: 10.1371/journal.pone.0023375

Figure Lengend Snippet: After 2, 3, 5, 7 and 10 days post seeding, mRNA was extracted and realtime PCR was performed using specific primers for connexin 43 (A) and N-cadherin (B). When samples were pre-coated with EMD, up to 4 fold increases in gene expression were observed for connexin 43 at 2, 3 and 5 days post seeding (A). 3 fold increases in gene expression of N-cadherin were also observed (B). Additional samples were extracted for western blot analysis (C). Elevated levels of both connexin 43 and N-cadherin were observed at 2, 3, 5 and 7 days post seeding. * denotes significant difference between EMD treated sample and respective control sample (p<0.05). Data shown is the average value from 3 independent experiments (3 replicates per experiment) ± SE.

Article Snippet: Cells were permealized with Triton X-100 for 5 minutes, followed by staining with antibodies against connexin 43 (sc-9059) or N-cadherin (sc-7939) (Santa Cruz Biotechnologies Inc., Heidelberg, Germany) antibodies, followed by a goat anti-rabbit IgG conjugated to texas red at a dilution of 1∶200 in 0.5% PBS/bovine serum albumin (Invitrogen).

Techniques: Expressing, Western Blot