Journal: PLoS ONE

Article Title: Antiproliferative Effects of DNA Methyltransferase 3B Depletion Are Not Associated with DNA Demethylation

doi: 10.1371/journal.pone.0036125

Figure Lengend Snippet: (A) Array Northern analysis of DNMT3B mRNA expression on Affymetrix HGU133Plus2.0 arrays in tumor (T) tissue samples compared to normal (N) tissues of the same organ. n indicates the number of analyzed samples for the specified tissue type. Data are shown as mean±SD; *P<0.05 relative to normal tissue (N), **P<0.01 relative to normal tissue (N), ***P<0.001 relative to normal tissue (N), determined by student's t-test. (B) Quantitative real-time PCR analysis of DNMT3B mRNA expression in colon cancer cell lines and normal cells (HMEC-T53, WI-38). Expression values are means of triplicates and were calculated relative to GAPDH expression. Error bars represent standard errors. (C) Immunoblot of DNMT3B protein expression in lysates from various colon cancer cell lines and normal cells (HMEC-T53, WI-38). ß-actin was used as a loading control.

Article Snippet: WI-38 cells were maintained in MEM Earle's medium supplemented with L-glutamine (Biochrom) and 10% fetal calf serum (PAA), and HMEC-T53 cells were grown in Mammary Epithelial Cell Growth Medium (Promocell: C-21010) and SupplementMix (Promocell: C-39115).

Techniques: Northern Blot, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Cx43 Promotes Endothelial Cell Migration and Angiogenesis via the Tyrosine Phosphatase SHP-2

doi: 10.3390/ijms23010294

Figure Lengend Snippet: Downregulation of Cx43 reduces endothelial cell migration. ( a ) Western blot analysis of Cx43, Cx40, and Cx37 revealed that Cx43 is expressed in both human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC), whereas Cx40 and Cx37 are not expressed in HMEC. Cx43 (Cx43), Cx40 (Cx), and Cx37 (Cx) expressing HeLa cells were used as the positive controls. CTL: Wildtype HeLa cell lysate. ( b ) Western blot analysis of Cx43 in HMEC transfected with 100 nM siRNA against Cx43 or non-silencing control siRNA (Ctrl) for 48 h. The Cx43 expression was analysed using a polyclonal Cx43 antibody. Immunostaining for GAPDH was used as the loading control. ( c ) Serum-induced cell migration of HMEC transfected with a Cx43-siRNA or a control siRNA (Ctrl) in response to 10% FCS was assessed in a wound assay. Downregulation of Cx43 significantly decreased cell migration, represented as accumulated distance (*** p < 0.001 Ctrl-siRNA vs. Cx43-siRNA, n = 6, 3 independent cell cultures). ( d ) Representative single cell traces of migrated HMEC.

Article Snippet: Human microvascular endothelial cells (HMEC) were provided by Ades et al. [ ] and were cultured in endothelial cell growth medium (DMEM supplemented with 10% fetal calf serum (FCS), 10% endothelial growth media (PromoCell, Heidelberg, Germany), and 1% penicillin/streptomycin (Sigma Aldrich)).

Techniques: Migration, Western Blot, Expressing, Transfection, Immunostaining