primary antibodies for mouse cx43 cx-1b1  (Thermo Fisher)

Journal: Prenatal Diagnosis

Article Title: Role of Myofibroblasts in the Repair of Iatrogenic Preterm Membranes Subjected to Mechanical Stimulation

doi: 10.1002/pd.6722

Figure Lengend Snippet: Myofibroblast morphology in human iatrogenic preterm AM. Fetal membranes were taken from a 31 year old patient who underwent open fetal surgery for correction of spina bifida neural tube defect that took place at 24 + 0 weeks gestational age leading to late trimester preterm delivery at 34 + 6 weeks (Case #1). Myofibroblast morphology was examined in αSMA expressing cells (green arrow) by IMF confocal microscopy in control AM specimens (A) and near the edge of the defect site (B, C). Cx43 was detected by immunostaining (pink arrow, C) and collagen by SHG imaging (red arrow). Control AM specimens were taken 5 cm away from the defect site. Signals for blue (DAPI), green (αSMA), pink (Cx43) and red (collagen) were detected by IMF confocal microscopy and SHG imaging. The dotted white lines show the length of the wound edge (WE) in the iatrogenic preterm AM specimen. Sale bar = 100 μM (A, B) and 300 μM (C). The white dotted box in (B) shows an enlarged image to show Cx43 localized in cell bodies.

Article Snippet: AM specimens were incubated with primary antibodies for mouse Cx43 (1:100, ThermoFisher Scientific, CX‐1B1) and rabbit αSMA (1:100, Abcam, ab5694) at 4°C overnight as described 44 Specimens were washed in PBS and incubated with secondary antibodies for Alexa Fluor 568 anti‐mouse or 488 anti‐rabbit at room temperature for 2 h (both 1:1000, ThermoFisher Scientific) and counterstained with 1 μg/mL DAPI for 20 min (1:1000) to detect nuclei.

Techniques: Expressing, Confocal Microscopy, Control, Immunostaining, Imaging

Journal: Prenatal Diagnosis

Article Title: Role of Myofibroblasts in the Repair of Iatrogenic Preterm Membranes Subjected to Mechanical Stimulation

doi: 10.1002/pd.6722

Figure Lengend Snippet: Cx43 plaque formation in human iatrogenic preterm ruptured amniotic membrane. Representative images obtained by confocal microscopy are shown in the epithelial (A) and fibroblast layer (B) of the preterm AM defect from one late third trimester iatrogenic preterm donor (Case #1). The distribution of Cx43 was analyzed per tissue area for comparisons between the epithelial and fibroblast layers (C) or per cell nucleus (D). Error bars represent the mean and SEM values of 6 replicates from three late trimester iatrogenic preterm donors (34 + 0 weeks to 35 + 0 weeks, Case #1–3). Significant differences are indicated by *** p < 0.001. Statistical comparisons are indicated for control AM specimens and AM defect ( p < 0.001***) in the epithelial layer ( p < 0.001 +++ ) and fibroblast layer ( p < 0.001 $$$ ). The dotted white and black lines show the length of the wound edge (WE) in the preterm AM specimen. Sale bar = 100 μM (A, B).

Article Snippet: AM specimens were incubated with primary antibodies for mouse Cx43 (1:100, ThermoFisher Scientific, CX‐1B1) and rabbit αSMA (1:100, Abcam, ab5694) at 4°C overnight as described 44 Specimens were washed in PBS and incubated with secondary antibodies for Alexa Fluor 568 anti‐mouse or 488 anti‐rabbit at room temperature for 2 h (both 1:1000, ThermoFisher Scientific) and counterstained with 1 μg/mL DAPI for 20 min (1:1000) to detect nuclei.

Techniques: Membrane, Confocal Microscopy, Control

Journal: Prenatal Diagnosis

Article Title: Role of Myofibroblasts in the Repair of Iatrogenic Preterm Membranes Subjected to Mechanical Stimulation

doi: 10.1002/pd.6722

Figure Lengend Snippet: Myofibroblast nuclei deformation in human iatrogenic preterm AM. AM specimens were immunostained for αSMA to detect myofibroblast (pink arrow) nuclei (blue arrow, A). Circularity values for the nuclei shape of myofibroblasts expressing αSMA in the epithelial layer were compared to the fibroblast layer in control specimens and AM defect (B). Nuclei values close to 1 represent a perfect circle in contrast to zero, which represents a more elongated shape. The total number of nuclei counted ranged from 1202 to 1582 from 6 specimens (Case #1–3). Signals for blue (DAPI), green (αSMA) and pink (Cx43) were detected by immunofluorescence confocal microscopy. The dotted white lines show the length of the wound edge (WE) in the iatrogenic preterm AM specimen. Scale bar = 100 μM. *** p < 0.001.

Article Snippet: AM specimens were incubated with primary antibodies for mouse Cx43 (1:100, ThermoFisher Scientific, CX‐1B1) and rabbit αSMA (1:100, Abcam, ab5694) at 4°C overnight as described 44 Specimens were washed in PBS and incubated with secondary antibodies for Alexa Fluor 568 anti‐mouse or 488 anti‐rabbit at room temperature for 2 h (both 1:1000, ThermoFisher Scientific) and counterstained with 1 μg/mL DAPI for 20 min (1:1000) to detect nuclei.

Techniques: Expressing, Control, Immunofluorescence, Confocal Microscopy

Journal: Prenatal Diagnosis

Article Title: Role of Myofibroblasts in the Repair of Iatrogenic Preterm Membranes Subjected to Mechanical Stimulation

doi: 10.1002/pd.6722

Figure Lengend Snippet: Effects of mechanical stimulation in human iatrogenic preterm AM. Preterm AM specimens were traumatized with a needed to create a 0.8‐mm defect and subjected to cyclic tensile strain (CTS) for 24 h. Mechanical stimulation was applied intermittently at 2% strain and 1 Hz frequency in the presence and absence of 50 μM Cx43 antisense (Cx43as). Absolute values for GAG (A), collagen (B) and elastin content (C) were normalized to dry tissue weight. PGE 2 release was quantified in media samples (D). Explants cultured without cyclic tensile strain (−CTS) were compared to +CTS specimens. Error bars represent the mean and SEM values of 24 replicates from three late third trimester iatrogenic preterm donors (34 + 0 weeks to 35 + 0 weeks, Case #1–3). Significant comparisons are indicated for −CTS and +CTS conditions where *** p < 0.001. All other comparisons (not indicated) were not significantly different.

Article Snippet: AM specimens were incubated with primary antibodies for mouse Cx43 (1:100, ThermoFisher Scientific, CX‐1B1) and rabbit αSMA (1:100, Abcam, ab5694) at 4°C overnight as described 44 Specimens were washed in PBS and incubated with secondary antibodies for Alexa Fluor 568 anti‐mouse or 488 anti‐rabbit at room temperature for 2 h (both 1:1000, ThermoFisher Scientific) and counterstained with 1 μg/mL DAPI for 20 min (1:1000) to detect nuclei.

Techniques: Cell Culture

Journal: Prenatal Diagnosis

Article Title: Role of Myofibroblasts in the Repair of Iatrogenic Preterm Membranes Subjected to Mechanical Stimulation

doi: 10.1002/pd.6722

Figure Lengend Snippet: Effects of mechanical stimulation on gene expression in human iatrogenic preterm AM. Preterm AM specimens were traumatized with a needed to create a 0.8‐mm defect and subjected to cyclic tensile strain (+CTS) for 4 and 24 h. Mechanical stimulation was applied intermittently at 2% strain and 1 Hz frequency in the presence and absence of 50 μM Cx43 antisense (Cx43as). Gene expression of Cx43 (A) and TGFβ (B) are presented as ratio values and normalized to control values. In all cases, error bars represent the mean and SEM values of 12 replicates from three late third trimester iatrogenic preterm donors (34 + 0 weeks to 35 + 0 weeks, Case #1–3). Significant comparisons are indicated for −CTS and +CTS conditions where *** p < 0.001. All other comparisons (not indicated) were not significantly different.

Article Snippet: AM specimens were incubated with primary antibodies for mouse Cx43 (1:100, ThermoFisher Scientific, CX‐1B1) and rabbit αSMA (1:100, Abcam, ab5694) at 4°C overnight as described 44 Specimens were washed in PBS and incubated with secondary antibodies for Alexa Fluor 568 anti‐mouse or 488 anti‐rabbit at room temperature for 2 h (both 1:1000, ThermoFisher Scientific) and counterstained with 1 μg/mL DAPI for 20 min (1:1000) to detect nuclei.

Techniques: Expressing, Control

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Cx43 hemichannels are involved in endothelial cell migration. a , Representative images of the endothelial cell migration observed in the wound-healing assay in control conditions. Primary cultures of mesenteric endothelial cells were prepared for wound-healing assay and cell migration was analyzed at 0, 10, 15 or 20 h after scratching the monolayer. Yellow lines are only intended to highlight the migration front and are not a reference for migration analysis. b , Quantitative analysis of the endothelial cells movement in control conditions into the cell-free scratched area at different time points (10, 15 and 20 h). c , Quantitative analysis of the endothelial cell migration observed before (Control) and after the treatment with 50 µM 18-ß-Glycyrrhetenic acid (ß-GA), a general Cx-formed channel inhibitor, or 300 µM TAT-Gap19 (Gap19), a specific Cx43 hemichannel blocking peptide. d , Representative images of immunofluorescence analysis of Cx43 expression in primary cultures of mesenteric endothelial cells in control conditions (monolayer) and in the migration front 4 h after scratching the monolayer. Cell nuclei are highlighted by the staining with DAPI (blue). Numbers inside the bars indicate the n value. Values are means ± SEM. *, P < 0.05 vs. Control by one-way ANOVA plus Bonferroni post hoc test

Article Snippet: Endothelial cells were fixed with 4% PFA, blocked with 3% BSA in PBS and incubated overnight at 4 °C with rabbit primary antibodies directed against cav-1 (1:100, Thermo Scientific, IL, USA), or SNO-cys (1:300, Sigma Aldrich, MO, USA) or a mouse primary antibody directed against Cx43 (1:100, BD-Transduction Labs, KY, USA), and then, with Alexa 568-labeled goat anti-rabbit or Alexa 488-labeled anti-mouse secondary antibodies (Molecular Probes, OR, USA) for 1 h at room temperature, as appropriate.

Techniques: Migration, Wound Healing Assay, Blocking Assay, Immunofluorescence, Expressing, Staining

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Cx43 hemichannels are not involved in endothelial cell proliferation. a , Analysis of endothelial cell proliferation by the bromodeoxyuridine (BrdU) incorporation assay in control conditions and after the treatment with the Cx43 hemichannel blocking peptide TAT-Gap19 (Gap19, 300 µM). Cell proliferation was evaluated in primary cultures of mesenteric endothelial cells at 40% or at 80% of confluence. b , Representative images of the immunofluorescence detection of BrdU incorporation into endothelial cells of the migration front and the monolayer in control conditions or in the presence of Gap19. c , Fluorescence intensity analysis of the experiments shown in b . BrdU+ denotes endothelial cells positive for BrdU. Numbers inside the bars indicate the n value. Values are means ± SEM

Article Snippet: Endothelial cells were fixed with 4% PFA, blocked with 3% BSA in PBS and incubated overnight at 4 °C with rabbit primary antibodies directed against cav-1 (1:100, Thermo Scientific, IL, USA), or SNO-cys (1:300, Sigma Aldrich, MO, USA) or a mouse primary antibody directed against Cx43 (1:100, BD-Transduction Labs, KY, USA), and then, with Alexa 568-labeled goat anti-rabbit or Alexa 488-labeled anti-mouse secondary antibodies (Molecular Probes, OR, USA) for 1 h at room temperature, as appropriate.

Techniques: BrdU Incorporation Assay, Blocking Assay, Immunofluorescence, Migration, Fluorescence

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Cx43-formed hemichannels, but not gap junction channels, are associated with endothelial cell migration. a , Representative images of dye coupling assay (left) and the analysis of the number of coupled cells via gap junction communication (right) attained in the intact monolayer and in the migration front after scratching the monolayer. Dye coupling was assessed by measuring after 2 min the diffusion to neighboring cells (coupled cells) of the ethidium bromide microinjected into a single endothelial cell. The yellow diamond indicates the microinjected cell. b , Representative images of the ethidium uptake observed in primary cultures of mesenteric endothelial cells in control conditions and in the presence of the Cx blocking peptide 37,43 Gap27 (200 µM) or the Cx43 hemichannel inhibitor TAT-Gap19 (Gap19, 300 µM) (left). Ethidium uptake was evaluated 15 min after scratching the monolayer and cells were incubated with the dye for 15 min, as shown in the time course of ethidium uptake observed in the intact monolayer and in the migration front (b, right top). Dot yellow lines depict the edge of the migration front. In addition, the analysis of the ethidium uptake rate achieved in the intact monolayer and in the migration front in control conditions and in the presence of 37,43 Gap27 or Gap19 is also shown (b, right bottom). The rate of ethidium uptake was assessed by calculating the slope of the increase in fluorescence intensity along the time. Changes in ethidium-fluorescence signal are expressed in arbitrary units (a.u.). Numbers inside the bars indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Monolayer by two-way ANOVA. ††, P < 0.01 and †††, P < 0.001 vs. Migration front in control conditions (Control) by one-way ANOVA plus Bonferroni post hoc test. &, P < 0.001 vs. Migration front by paired Student’s t-test

Article Snippet: Endothelial cells were fixed with 4% PFA, blocked with 3% BSA in PBS and incubated overnight at 4 °C with rabbit primary antibodies directed against cav-1 (1:100, Thermo Scientific, IL, USA), or SNO-cys (1:300, Sigma Aldrich, MO, USA) or a mouse primary antibody directed against Cx43 (1:100, BD-Transduction Labs, KY, USA), and then, with Alexa 568-labeled goat anti-rabbit or Alexa 488-labeled anti-mouse secondary antibodies (Molecular Probes, OR, USA) for 1 h at room temperature, as appropriate.

Techniques: Migration, Diffusion-based Assay, Blocking Assay, Incubation, Fluorescence

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Endothelial cell migration depends on a Cx43-formed channel-mediated increase in intracellular Ca 2+ concentration ([Ca 2+ ] i ). a , Representative images (left) and fluorescence intensity analysis (right) of the increase in [Ca 2+ ] i observed in endothelial cells of the migration front 15 min after scratching the monolayer in control conditions and in the presence of 50 µM 18-β-Glycyrrhetenic acid (ß-GA), a general Cx-formed channel blocker, or 200 µM 37,43 Gap27, a peptide designed to block channels formed by Cx37 or Cx43. Variations in the levels of [Ca 2+ ] i were assessed with the fluorescent Ca 2+ indicator Fluo-4. b , Representative images of the changes in [Ca 2+ ] i of endothelial cells (left) in which the differences in the subcellular distribution of the Ca 2+ signal attained in a cell of the migration front (Cell 2) and a cell within the monolayer (Cell 1) are highlighted in a 3D analysis (middle and right). c , Fluorescence intensity analysis of the Fluo-4 signal measured along the endothelial cells length (from back to front) in the migration front and the monolayer. d , Analysis of the changes in [Ca 2+ ] i levels attained in the rear and anterior edge of endothelial cells of the migration front in control conditions and after the treatment with ß-GA or 37,43 Gap27. Changes in Fluo-4 signal are expressed as the area under the curve (AUC). Note that cells were treated with the Cx blocking peptide 37,43 Gap27 for only 10 min to inhibit Cx hemichannels, without affecting gap junction channels. Dot red lines depict the edge of the migration front. Numbers inside the bars or in parentheses indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Control by one-way ANOVA plus Bonferroni post hoc test. †††, P < 0.001 vs. Anterior by paired Student’s t-test

Article Snippet: Endothelial cells were fixed with 4% PFA, blocked with 3% BSA in PBS and incubated overnight at 4 °C with rabbit primary antibodies directed against cav-1 (1:100, Thermo Scientific, IL, USA), or SNO-cys (1:300, Sigma Aldrich, MO, USA) or a mouse primary antibody directed against Cx43 (1:100, BD-Transduction Labs, KY, USA), and then, with Alexa 568-labeled goat anti-rabbit or Alexa 488-labeled anti-mouse secondary antibodies (Molecular Probes, OR, USA) for 1 h at room temperature, as appropriate.

Techniques: Migration, Concentration Assay, Fluorescence, Blocking Assay

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Cx43 hemichannel activation leads to subcellular re-distribution of caveolin-1 (Cav-1) and Cx43 in migrating endothelial cells. a , Immunofluorescence analysis of the subcellular distribution of Cav-1 (red) and Cx43 (green) in control conditions and after the treatment with 300 µM TAT-Gap19 (Gap19), a specific blocker of Cx43 hemichannels. Cell nuclei are highlighted by the staining with DAPI (blue). The square indicatres the area that was enlarged on the right. b and c , Densitometric analysis of Cx43 ( b ) and Cav-1 ( c ) signal in the rear and anterior part of endothelial cells of the migration front in control conditions and in the presence of Gap19. Numbers inside the bars indicate the n value. Values are means ± SEM. **, P < 0.01 vs. Anterior by paired Student’s t-test

Article Snippet: Endothelial cells were fixed with 4% PFA, blocked with 3% BSA in PBS and incubated overnight at 4 °C with rabbit primary antibodies directed against cav-1 (1:100, Thermo Scientific, IL, USA), or SNO-cys (1:300, Sigma Aldrich, MO, USA) or a mouse primary antibody directed against Cx43 (1:100, BD-Transduction Labs, KY, USA), and then, with Alexa 568-labeled goat anti-rabbit or Alexa 488-labeled anti-mouse secondary antibodies (Molecular Probes, OR, USA) for 1 h at room temperature, as appropriate.

Techniques: Activation Assay, Immunofluorescence, Staining, Migration

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Activation of Cx43-formed hemichannels is involved in the progress of angiogenesis in vitro. Representative images of the formation of tubular-like structures in Matrigel by primary cultures of mesenteric endothelial cells after 12 h in control conditions or in the presence of 300 µM TAT-Gap19 (Gap19), a specific blocker of Cx43 hemichannels (left). In addition, the analysis of the development of tubular-like structure formation by the calculation of the angiogenic index observed after 6 and 12 h in control conditions and in the presence of Gap19 is also shown (right). Values are means ± SEM. *, P < 0.05 vs. Control by unpaired Student’s t-test

Article Snippet: Endothelial cells were fixed with 4% PFA, blocked with 3% BSA in PBS and incubated overnight at 4 °C with rabbit primary antibodies directed against cav-1 (1:100, Thermo Scientific, IL, USA), or SNO-cys (1:300, Sigma Aldrich, MO, USA) or a mouse primary antibody directed against Cx43 (1:100, BD-Transduction Labs, KY, USA), and then, with Alexa 568-labeled goat anti-rabbit or Alexa 488-labeled anti-mouse secondary antibodies (Molecular Probes, OR, USA) for 1 h at room temperature, as appropriate.

Techniques: Activation Assay, In Vitro

Journal: Biology Direct

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

doi: 10.1186/s13062-023-00408-3

Figure Lengend Snippet: Activation of Cx43-formed hemichannels in endothelial cells of migration front depends on NO-mediated S-nitrosylation. a , Representative images of ethidium uptake in primary cultures of mesenteric endothelial cells (left) and the analysis of ethidium uptake rate observed in the migration front and in the monolayer in control conditions and after the treatment with 100 µM N G -nitro-L-arginine (L-NA) or 50 µM ascorbic acid (AA). The rate of ethidium uptake was assessed by calculating the slope of the increase in fluorescence intensity along the time. b , Representative images (left) and fluorescence intensity analysis (right) of the increase in [Ca 2+ ] i observed in endothelial cells of the migration front in control conditions and in the presence of 100 µM L-NA or 50 µM AA. Variations in the levels of [Ca 2+ ] i were assessed with the fluorescent Ca 2+ indicator Fluo-4. c , Representative images (left) and quantitative analysis (right) of the endothelial cell migration observed in the wound-healing assay just after scratching the monolayer (0 h) and after 15 h in control conditions and in the presence of L-NA or AA. Yellow lines are only intended to highlight the migration front and are not a reference for migration analysis. d , Detection of total protein S-nitrosylation by immunofluorescence (left) and densitometric analysis of the fluorescence intensity (right) observed in the migration front and the monolayer of primary cultures of endothelial cells in control conditions and after the treatment with L-NA or AA. Cell nuclei are highlighted by the staining with DAPI (blue). Numbers inside the bars indicate the n value. Values are means ± SEM. *, P < 0.05 and **, P < 0.01 vs. Control by one-way ANOVA plus Bonferroni post hoc test

Article Snippet: Endothelial cells were fixed with 4% PFA, blocked with 3% BSA in PBS and incubated overnight at 4 °C with rabbit primary antibodies directed against cav-1 (1:100, Thermo Scientific, IL, USA), or SNO-cys (1:300, Sigma Aldrich, MO, USA) or a mouse primary antibody directed against Cx43 (1:100, BD-Transduction Labs, KY, USA), and then, with Alexa 568-labeled goat anti-rabbit or Alexa 488-labeled anti-mouse secondary antibodies (Molecular Probes, OR, USA) for 1 h at room temperature, as appropriate.

Techniques: Activation Assay, Migration, Fluorescence, Wound Healing Assay, Immunofluorescence, Staining

Journal: Heliyon

Article Title: Differentiation of cardiomyocyte-like cells from human amniotic fluid mesenchymal stem cells by combined induction with human platelet lysate and 5-azacytidine

doi: 10.1016/j.heliyon.2020.e04844

Figure Lengend Snippet: The primers for RT–qPCR and products size.

Article Snippet: After fixation, the cells were blocked in 10% AB-serum in 1% BSA-PBS for 30 min at room temperature, and then incubated with mouse anti-human Cx43 primary antibodies (Sigma-aldrich, USA) for 2 h at 37 °C.

Techniques:

Journal: Heliyon

Article Title: Differentiation of cardiomyocyte-like cells from human amniotic fluid mesenchymal stem cells by combined induction with human platelet lysate and 5-azacytidine

doi: 10.1016/j.heliyon.2020.e04844

Figure Lengend Snippet: A: hPL ability to up-regulate cardiomyogenic specific genes expression. Each graph displays the expression levels of GATA4 , cTnT , Cx4 3 and Nkx2.5 , which were normalized to GAPDH and were relative to the control group. Data are presented as mean ± S.E. values. ∗ statistically significant versus control. # statistically significant between group. B: hPL combined with 5-aza ability to up-regulate cardiomyogenic specific genes expression. Each graph displays the expression levels of GATA4 , cTnT , Cx43 and Nkx2.5 , which were normalized to GAPDH and were relative to the control group. Data are presented as mean ± S.E. values. ∗ statistically significant versus control. # statistically significant between group.

Article Snippet: After fixation, the cells were blocked in 10% AB-serum in 1% BSA-PBS for 30 min at room temperature, and then incubated with mouse anti-human Cx43 primary antibodies (Sigma-aldrich, USA) for 2 h at 37 °C.

Techniques: Expressing

Journal: Heliyon

Article Title: Differentiation of cardiomyocyte-like cells from human amniotic fluid mesenchymal stem cells by combined induction with human platelet lysate and 5-azacytidine

doi: 10.1016/j.heliyon.2020.e04844

Figure Lengend Snippet: Ability of hPL to promote the differentiation of hAF-MSCs into cardiomyocyte-like cells. Each graph displays the expression levels of GATA4 , cTnT , Cx43 and Nkx2. 5 that were normalized to GAPDH and were relative to the control group. Data are presented as mean ± S.E. values. ∗ statistically significant versus control. # statistically significant between group.

Article Snippet: After fixation, the cells were blocked in 10% AB-serum in 1% BSA-PBS for 30 min at room temperature, and then incubated with mouse anti-human Cx43 primary antibodies (Sigma-aldrich, USA) for 2 h at 37 °C.

Techniques: Expressing

Journal: Heliyon

Article Title: Differentiation of cardiomyocyte-like cells from human amniotic fluid mesenchymal stem cells by combined induction with human platelet lysate and 5-azacytidine

doi: 10.1016/j.heliyon.2020.e04844

Figure Lengend Snippet: Detection of cardiomyogenic specific proteins; immunofluorescence staining FITC (495 nm/519 nm), green color for all cardiomyogenic specific proteins (A–I) and immunoenzymatic staining (J–L); GATA4 (localized in nucleus) staining (A) control group, (B) 10 μM 5-aza induced group, (C) 10 μM 5-aza with 20% hPL induced group; cTnT (localized in cytoplasm) staining (D) control group, (E) 10 μM 5-aza induced group, (F) 10 μM 5-aza 20% hPL induced group; Nkx2.5 (localized in nucleus) staining (G) control group, (H) 10 μM 5-aza induced group, (I) 10 μM 5-aza with 20% hPL induced group; Cx43 (localized in cell membrane) staining (J) control group, (K) 10 μM 5-aza induced group (black arrow), (L) 10 μM 5-aza with 20% hPL induced group (black arrow). (A-C and G-I) insets without nuclear counterstain showing no nuclear staining of two key core cardiac transcription factors. Scale bar = 100 μm.

Article Snippet: After fixation, the cells were blocked in 10% AB-serum in 1% BSA-PBS for 30 min at room temperature, and then incubated with mouse anti-human Cx43 primary antibodies (Sigma-aldrich, USA) for 2 h at 37 °C.

Techniques: Immunofluorescence, Staining

Journal: Heliyon

Article Title: Differentiation of cardiomyocyte-like cells from human amniotic fluid mesenchymal stem cells by combined induction with human platelet lysate and 5-azacytidine

doi: 10.1016/j.heliyon.2020.e04844

Figure Lengend Snippet: Image J analysis showing the results of CTCF (the expression levels of GATA4, cTnT, Nkx2.5 and Cx43 proteins signal). Data are presented as mean ± S.E. values. ∗ statistically significant versus control.

Article Snippet: After fixation, the cells were blocked in 10% AB-serum in 1% BSA-PBS for 30 min at room temperature, and then incubated with mouse anti-human Cx43 primary antibodies (Sigma-aldrich, USA) for 2 h at 37 °C.

Techniques: Expressing

Journal: Medicine

Article Title: Effect of gap junctions on RAW264.7 macrophages infected with H37Rv

doi: 10.1097/MD.0000000000012125

Figure Lengend Snippet: H37Rv infection induces host macrophage connexin expression. RAW264.7 cells were infected by H37Rv strains for 12 hours. Total RNA and protein were tested using RT-PCR (Cx37 and Cx43 mRNA) (A) and Western blotting (Cx43 protein) (B). Both the control and infection cells were collected and tested using immunofluorescence analysis (C). Data are expressed as mean±SD from 3 independent experiments. ∗ P < .05 versus control; ∗∗ P < .01 versus control.

Article Snippet: [ ] A concentration of rabbit anti-mouse Cx43 primary antibodies (1:1000) was purchased from Cell Signaling Technology (Danvers, MA), and mouse anti-mouse β-actin (1:1000) was purchased from ZSGB-BIO (Beijing, China).

Techniques: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Immunofluorescence