Journal: Molecules

Article Title: Impaired Expression of Cytoplasmic Actins Leads to Chromosomal Instability of MDA-MB-231 Basal-Like Mammary Gland Cancer Cell Line

doi: 10.3390/molecules26082151

Figure Lengend Snippet: Figure 1. Downregulation of β- or γ-actin expression by corresponding shRNAs in MDA-MB-231 breast cancer cells leads to phenotype changes. (A). Immunofluorescent staining of MDA-MB-231 cells with β- or γ-actin downregulation by corresponding shRNAs. Arrowheads mark leading edge with actin enrichment. Bar, 10 µm. (B). Downregulation of cytoplasmic β- or γ-actin in MDA-MB-231 cells. WB analysis. Graphs represent relative actins expression (Mean ± SEM). For cells with shRNA compared with control (β-actin): p = 0.0015 (shRNA to β-actin); p = 0.007 (shRNA to γ-actin); (γ-actin): p = 0.0037 (shRNA to β-actin); p = 0.0048 (shRNA to γ-actin); Mann–Whitney U test, n = 3. Values of p < 0.01 (**) were considered as statistically significant.

Article Snippet: Mouse monoclonal antibodies to: β-actin (MCA5775GA, AbD Serotec); γ-actin (MCA5776GA, AbD Serotec, Raleigh, NC, USA); pan actin (4968, Cell Signaling) were used.

Techniques: Expressing, Staining, shRNA, Control, MANN-WHITNEY

Journal: Molecules

Article Title: Impaired Expression of Cytoplasmic Actins Leads to Chromosomal Instability of MDA-MB-231 Basal-Like Mammary Gland Cancer Cell Line

doi: 10.3390/molecules26082151

Figure Lengend Snippet: Figure 2. Downregulation of β- or γ-actin expression by corresponding shRNAs in MDA-MB-231 breast cancer cells leads to cell cycle changes. (A). After 6d of β- or γ-actin depletion by corresponding shRNAs MDA-MB-231 cells were stained with propidium iodide. High content analysis of cell cycle distributions of flow cytometric data was performed after 6d of β- or γ-actin depletion by corresponding shRNAs in MDA-MB-231 cells. (B). The effects of β- or γ-actin depletion on quantitative ratio of cells in interphase and different mitotic phases were analyzed by IF cytometry. The Y-axis indicates the number of cells in specific phases. Student’s t-test, n = 3. Values of p < 0.01 (**), and p < 0.05 (*) were considered as statistically significant. (C). The effects of β- or γ-actin depletion on mitotic phases were analyzed by IF cytometry. The pie chart shows the proportion of cells in different phases of mitosis.

Article Snippet: Mouse monoclonal antibodies to: β-actin (MCA5775GA, AbD Serotec); γ-actin (MCA5776GA, AbD Serotec, Raleigh, NC, USA); pan actin (4968, Cell Signaling) were used.

Techniques: Expressing, Staining, High Content Screening, Cytometry

Journal: Molecules

Article Title: Impaired Expression of Cytoplasmic Actins Leads to Chromosomal Instability of MDA-MB-231 Basal-Like Mammary Gland Cancer Cell Line

doi: 10.3390/molecules26082151

Figure Lengend Snippet: Figure 3. Karyotypic analysis of MDA-MB-231 breast cancer cells. (A). A representative G-banded karyotype of MDA- MB-231 cell line. (B). Composite G-banded karyotype of the near-triploid cell line MDA-MB-231, showing structural and numerical changes. Arrows point to main chromosomal alterations. Mar—Marker chromosome. (C,D,F,G). The representa- tive chromosome aberrations in β-actin (C,D) and γ-actin (F,G) -depleted MDA-MB-231 cells: dicentric chromosomes (C,F) and acentric fragment (C), ring chromosomes (D,G). Arrows point to aberrations. E, H. Endoreduplication in β-actin (E) and γ-actin (H) -depleted MDA-MB-231 cell.

Article Snippet: Mouse monoclonal antibodies to: β-actin (MCA5775GA, AbD Serotec); γ-actin (MCA5776GA, AbD Serotec, Raleigh, NC, USA); pan actin (4968, Cell Signaling) were used.

Techniques: Marker

Journal: BMC Endocrine Disorders

Article Title: A plausible role for actin gamma smooth muscle 2 ( ACTG2 ) in small intestinal neuroendocrine tumorigenesis

doi: 10.1186/s12902-016-0100-3

Figure Lengend Snippet: Analysis of ACTG2 protein expression in SI-NETs by immunohistochemistry ( a - d ) using ACTG2 antibody (NB100-91649 Novus Biologicals) and western blotting ( e ) using another ACTG2 antibody (TA313418 Origene). a Negatively stained tumor cells and strong stromal staining (20x). b Areas with positively stained tumor cells, and negative stromal staining (20x). c Weak staining in all tumor cells (20x). d No staining in absence of primary antibody (20x). e Western blotting analysis showing antibody specificity and correlation to immunohistochemistry analysis. One band only was visualized in two tumors (lanes 2 and 3) displaying strong stromal staining, and no band was detected in two tumors (lanes 1 and 4) with no staining in both tumor and stromal cells. Lane 1, lymph node metastasis; lanes 2–4, primary tumors. Coomassie blue staining was used as loading control, ladder in kDa

Article Snippet: The tissues were treated with normal serum from goat (S-1000, Vector) and two different rabbit polyclonal anti-ACTG2 antibodies (NB100-91649 Novus Biologicals, diluted 1/200, and TA313418 Origene, diluted 1/80) and anti-chromogranin A antibodies (Ab-1, LK2H10, NeoMarkers, diluted 1/1000) were used and incubated.

Techniques: Expressing, Immunohistochemistry, Western Blot, Staining, Control

Journal: BMC Endocrine Disorders

Article Title: A plausible role for actin gamma smooth muscle 2 ( ACTG2 ) in small intestinal neuroendocrine tumorigenesis

doi: 10.1186/s12902-016-0100-3

Figure Lengend Snippet: Double immunofluorescence staining of intestinal mucosa. Chromogranin A is visualized as green, showing positively stained enterochromaffin cells. ACTG2 is visualized as red and no staining is detected in chromogranin A positive cells (yellow)

Article Snippet: The tissues were treated with normal serum from goat (S-1000, Vector) and two different rabbit polyclonal anti-ACTG2 antibodies (NB100-91649 Novus Biologicals, diluted 1/200, and TA313418 Origene, diluted 1/80) and anti-chromogranin A antibodies (Ab-1, LK2H10, NeoMarkers, diluted 1/1000) were used and incubated.

Techniques: Double Immunofluorescence Staining, Staining

Journal: BMC Endocrine Disorders

Article Title: A plausible role for actin gamma smooth muscle 2 ( ACTG2 ) in small intestinal neuroendocrine tumorigenesis

doi: 10.1186/s12902-016-0100-3

Figure Lengend Snippet: Effects on ACTG2 mRNA expression in CNDT2.5 cells after DZNep (3-deazaneplanocin A) and 1.0 μM EPZ-6438 treatment, a and b respectively

Article Snippet: The tissues were treated with normal serum from goat (S-1000, Vector) and two different rabbit polyclonal anti-ACTG2 antibodies (NB100-91649 Novus Biologicals, diluted 1/200, and TA313418 Origene, diluted 1/80) and anti-chromogranin A antibodies (Ab-1, LK2H10, NeoMarkers, diluted 1/1000) were used and incubated.

Techniques: Expressing

Journal: BMC Endocrine Disorders

Article Title: A plausible role for actin gamma smooth muscle 2 ( ACTG2 ) in small intestinal neuroendocrine tumorigenesis

doi: 10.1186/s12902-016-0100-3

Figure Lengend Snippet: a Effects on ACTG2 mRNA expression in CNDT2.5 cells after miR-145 transfection. b Effects on miR-145 expression after DZNep treatment. c Quantitative determination of cytoplasmic histone-associated-DNA-fragments (mono- and oligonucleosomes) after miR-145 transfection. Camptothecin at 0.1 μg/ml was used as positive control

Article Snippet: The tissues were treated with normal serum from goat (S-1000, Vector) and two different rabbit polyclonal anti-ACTG2 antibodies (NB100-91649 Novus Biologicals, diluted 1/200, and TA313418 Origene, diluted 1/80) and anti-chromogranin A antibodies (Ab-1, LK2H10, NeoMarkers, diluted 1/1000) were used and incubated.

Techniques: Expressing, Transfection, Positive Control

Journal: BMC Endocrine Disorders

Article Title: A plausible role for actin gamma smooth muscle 2 ( ACTG2 ) in small intestinal neuroendocrine tumorigenesis

doi: 10.1186/s12902-016-0100-3

Figure Lengend Snippet: a miR-145 expression levels in 24 SI-NETs, and in CNDT2.5. A significant ( p < 0.01) difference between primary tumors and liver metastases, and also between lymph node and liver metastasis ( p < 0.001) was observed. A tendency towards decreased expression in lymph node metastases compared to primary tumors was detected ( p = 0.09). b ACTG2 mRNA expression levels in 18 PT and 16 LNM. A significant ( p < 0.01) difference between primary tumors and lymph node metastases was observed. PT, primary tumor. LNM, lymph node metastasis. LM, liver metastasis

Article Snippet: The tissues were treated with normal serum from goat (S-1000, Vector) and two different rabbit polyclonal anti-ACTG2 antibodies (NB100-91649 Novus Biologicals, diluted 1/200, and TA313418 Origene, diluted 1/80) and anti-chromogranin A antibodies (Ab-1, LK2H10, NeoMarkers, diluted 1/1000) were used and incubated.

Techniques: Expressing

Journal: BMC Endocrine Disorders

Article Title: A plausible role for actin gamma smooth muscle 2 ( ACTG2 ) in small intestinal neuroendocrine tumorigenesis

doi: 10.1186/s12902-016-0100-3

Figure Lengend Snippet: a Colony formation assay in CNDT2.5 cells stably transfected with a plasmid expressing ACTG2 or with empty expression vector. b Western blotting demonstrating successful transfection of the DDK epitope fused to ACTG2. c Viability assay using WST-1 after transient overexpression of ACTG2. d Western blotting demonstrating successful transfection of the DDK epitope fused to ACTG2

Article Snippet: The tissues were treated with normal serum from goat (S-1000, Vector) and two different rabbit polyclonal anti-ACTG2 antibodies (NB100-91649 Novus Biologicals, diluted 1/200, and TA313418 Origene, diluted 1/80) and anti-chromogranin A antibodies (Ab-1, LK2H10, NeoMarkers, diluted 1/1000) were used and incubated.

Techniques: Colony Assay, Stable Transfection, Transfection, Plasmid Preparation, Expressing, Western Blot, Viability Assay, Over Expression

Journal: Biology of Reproduction

Article Title: Progesterone-Regulated Caspase 3 Action in the Mouse May Play a Role in Uterine Quiescence During Pregnancy Through Fragmentation of Uterine Myocyte Contractile Proteins

doi: 10.1095/biolreprod.108.070425

Figure Lengend Snippet: The effect of gestational age on the levels of full-length versus caspase 3-cleaved (A) smooth muscle α-actin and (B) smooth muscle γ-actin in the pregnant mouse uterus. A representative Western blot of the full-length smooth muscle α- and γ-actins and their caspase 3-cleaved fragments are shown. Smooth muscle α- and γ-actin full length and caspase 3-cleaved levels were normalized to calponin. Data are presented as mean ± SEM. *ROD for cleaved and full-length smooth muscle α- and γ-actin is significantly different (P < 0.005).

Article Snippet: Blots were probed using primary antibodies against calponin (1:250 dilution; EP798Y; Novus Biologicals, Littleton, CO), anti-caspase 3 cleaved form (1:1000 dilution; AB3623; Chemicon, Billerica, MA), smooth muscle α-actin (1:1000 dilution; CBL171; Chemicon), and smooth muscle γ-actin (1:1000 dilution; ACTG2; Novus Biologicals).

Techniques: Western Blot

Journal: Biology of Reproduction

Article Title: Progesterone-Regulated Caspase 3 Action in the Mouse May Play a Role in Uterine Quiescence During Pregnancy Through Fragmentation of Uterine Myocyte Contractile Proteins

doi: 10.1095/biolreprod.108.070425

Figure Lengend Snippet: The effect of gestational age on the mRNA expression levels of (A) smooth muscle α-actin and (B) smooth muscle γ-actin in the pregnant mouse uterus. Data are presented as mean ± SEM. *Significantly different (P < 0.005).

Article Snippet: Blots were probed using primary antibodies against calponin (1:250 dilution; EP798Y; Novus Biologicals, Littleton, CO), anti-caspase 3 cleaved form (1:1000 dilution; AB3623; Chemicon, Billerica, MA), smooth muscle α-actin (1:1000 dilution; CBL171; Chemicon), and smooth muscle γ-actin (1:1000 dilution; ACTG2; Novus Biologicals).

Techniques: Expressing

Journal: Biology of Reproduction

Article Title: Progesterone-Regulated Caspase 3 Action in the Mouse May Play a Role in Uterine Quiescence During Pregnancy Through Fragmentation of Uterine Myocyte Contractile Proteins

doi: 10.1095/biolreprod.108.070425

Figure Lengend Snippet: The effect of exogenous progesterone treatment on the gestational profile of full-length versus caspase 3-cleaved (A) smooth muscle α-actin and (B) smooth muscle γ-actin in the pregnant mouse uterus. Representative Western blots of the full length smooth muscle α- and γ-actins and their caspase 3-cleaved fragments are shown. The levels of smooth muscle α- and γ-actin were normalized to calponin. Data are presented as mean ± SD ROD for smooth muscle α- and γ-actin caspase 3-cleaved fragments. *Control samples significantly different from progesterone-treated samples (P < 0.05).

Article Snippet: Blots were probed using primary antibodies against calponin (1:250 dilution; EP798Y; Novus Biologicals, Littleton, CO), anti-caspase 3 cleaved form (1:1000 dilution; AB3623; Chemicon, Billerica, MA), smooth muscle α-actin (1:1000 dilution; CBL171; Chemicon), and smooth muscle γ-actin (1:1000 dilution; ACTG2; Novus Biologicals).

Techniques: Western Blot