Journal:

Article Title: Reduction of electrical coupling between microvascular endothelial cells by NO depends on connexin37

doi: 10.1152/ajpheart.01148.2008

Figure Lengend Snippet: Effect of DETA exposure on intercellular resistance in wild-type, connexin (Cx) 37 null, Cx40 null, and Cx43G60S (nonfunctional mutant) MMEC. DETA (500 μM, 3 h) increased resistance in cells derived from wild-type, Cx40 null, and Cx43G60S mice, but not in cells from Cx37 null mice. *Significant difference from the appropriate control group, P < 0.05; n = 16 in group 1 (pooled from all control experiments), n = 21 in group 2 (pooled), and n = 4 in groups 3–8.

Article Snippet: Peroxidase-labeled anti-rabbit IgG antibodies for anti-Cx37, anti-PKG, and anti-phosphoserine antibodies were purchased from Biolynx (Brockville, ON) and Cell Signaling Technology (Beverly, MA).

Techniques: Mutagenesis, Derivative Assay

Journal:

Article Title: Reduction of electrical coupling between microvascular endothelial cells by NO depends on connexin37

doi: 10.1152/ajpheart.01148.2008

Figure Lengend Snippet: Lack of effect of DETA on Cx37, Cx40, and Cx43 protein expression in wild-type MMEC. A: Cx37, Cx40, Cx43, and β-actin immunoblots from control and DETA-treated MMEC (500 μM, 3 h). B: densitometric ratios of Cx37, Cx40, and Cx43 to β-actin immunoblots for control and DETA-treated MMEC. DETA had no effect on Cx37, Cx40, or Cx43 protein expression; n = 3 in each group.

Article Snippet: Peroxidase-labeled anti-rabbit IgG antibodies for anti-Cx37, anti-PKG, and anti-phosphoserine antibodies were purchased from Biolynx (Brockville, ON) and Cell Signaling Technology (Beverly, MA).

Techniques: Expressing, Western Blot

Journal:

Article Title: Reduction of electrical coupling between microvascular endothelial cells by NO depends on connexin37

doi: 10.1152/ajpheart.01148.2008

Figure Lengend Snippet: Lack of effect of DETA on distribution of Cx37, Cx40, and Cx43 within wild-type MMEC. Top: representative examples of control (n = 9) and DETA-treated cells (500 μM, 3 h, n = 6) probed for Cx37. Although most of the immunoreactivity is cytoplasmic, occasional punctate staining (arrows) indicates putative Cx37 gap junctions. Comparable staining was also seen with another anti-Cx37 antibody (i.e., that used for immunoblotting) (data not shown). The superimposed oval uniform gray shapes are cell nuclei identified with Hoechst 33258 stain. Middle: representative examples of control (n = 7) and DETA-treated (n = 7) cells probed for Cx40. Numerous small Cx40 gap junctions are indicated by the punctate staining. The superimposed oval gray shapes are cell nuclei. Bottom: representative examples of control (n = 3) and DETA-treated (n = 3) cells probed for Cx43. Numerous Cx43 gap junctions of various sizes are evident, larger than those containing Cx37 or Cx40, whereas gray ovals indicate cell nuclei. Control experiments where the connexin primary antibodies were omitted showed only gray oval nuclei (data not shown). Bar at bottom right indicates 20-μm scale for all panels.

Article Snippet: Peroxidase-labeled anti-rabbit IgG antibodies for anti-Cx37, anti-PKG, and anti-phosphoserine antibodies were purchased from Biolynx (Brockville, ON) and Cell Signaling Technology (Beverly, MA).

Techniques: Staining, Western Blot

Journal: Cell Death & Disease

Article Title: Gap junctional communication promotes apoptosis in a connexin-type-dependent manner

doi: 10.1038/cddis.2013.105

Figure Lengend Snippet: The rate of apoptosis is dependent on the Cx type expressed and correlates to the GJ permeability. ( a ) Western blot analysis for the expression of Cx37, Cx40 and Cx43 in stably transfected HeLa cells compared with empty vector-transfected HeLa (CTL). ( b ) Immunofluorescence stainings of Cx proteins in stably transfected HeLa cells demonstrate typical localisation at the membrane of adjacent cells. HeLa-CTL cells were used as controls and showed no Cx43 expression by staining for Cx43. ( c ) Apoptosis (induced by 1 μ M SN, 3 h) of stably transfected HeLa cells is dependent on the Cx type expressed and significantly increased by expression of Cx40 or Cx43 compared with HeLa-CTL. Apoptosis was determined by annexin V-FITC/PI labelling and fold increase of apoptotic cells is expressed as mean±S.E.M.; * P <0.05 versus CTL; # P <0.05 versus Cx37; n ≥5 in at least three different cell cultures. ( d ) Gap junctional cell coupling of HeLa cells varies with the Cx type expressed and correlates with the rate of apoptosis. Cell coupling was measured by dye spreading (Alexa Fluor 488). The number of dye-stained cells is displayed as mean±S.E.M. ( n ≥18 dye injections in ≥3 different cell cultures; * P <0.05 versus CTL; # P <0.05 versus Cx37; NG). GAPDH, glyceraldehyde 3-phosphate dehydrogenase

Article Snippet: After rinsing with PBS, cells were permeabilised with 0.1% Triton X-100 (AppliChem) in PBS for 4 min. Unspecific antibody binding was blocked with PBS containing 0.5% BSA for 1 h. Cells were incubated with rabbit α -Cx37 (Alpha Diagnostic; 1 : 100), rabbit α -Cx40 (Alpha Diagnostic; 1 : 100) or rabbit α -Cx43 (Sigma Aldrich; 1 : 100) overnight at 4 °C.

Techniques: Permeability, Western Blot, Expressing, Stable Transfection, Transfection, Plasmid Preparation, Immunofluorescence, Membrane, Staining

Journal: Cancer Science

Article Title: Pattern of cell‐to‐cell transfer of micro RNA by gap junction and its effect on the proliferation of glioma cells

doi: 10.1111/cas.14029

Figure Lengend Snippet: Ability of gap junctions composed of connexin 32 (Cx32) or connexin 37 (Cx37) to transfer microRNAs (mi RNA s) between cervical cancer cells. A, Expression of Cx32 in HeLa‐Cx32 cells after 48 h of doxycycline (Dox) treatment. B, Function of gap junctions in HeLa‐Cx32 cells with induction of Cx32 expression. Scale bar, 10 μm. C, Expression of mi RNA s in receiving cells was analyzed by qPCR . D, Expression of Cx37 in HeLa cells with Cx37 plasmid stable transfection. GFP was labeled in HeLa cells as the coculture receiving cells. E, Function of gap junctions in HeLa cells transfected with Cx37 plasmid. Scale bar, 10 μm. F, Expression of mi RNA s was not significantly changed before and after the coculture. Columns represent the means of three experiments; bars represent the SEM . ** P < .01

Article Snippet: To overexpress Cx37, the Cx37 plasmid and the control plasmid (Cyagen Biosciences Inc.) were respectively transfected into HeLa cells.

Techniques: Expressing, Plasmid Preparation, Stable Transfection, Labeling, Transfection

Journal: MycoKeys

Article Title: Phylogenetic and morphological analyses of Coniochaeta isolates recovered from Inner Mongolia and Yunnan revealed three new endolichenic fungal species

doi: 10.3897/mycokeys.83.71140

Figure Lengend Snippet: GenBank accession numbers Coniochaeta species used for the phylogenetic analyses. T = ex-type isolates.

Article Snippet: In the tree constructed using the concatenated dataset, isolate CX37 (Taxon 2) formed a monophyletic clade with C. acaciae with high statistical support.

Techniques: