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: Cardiovascular research

Article Title: Functional tissue-engineered blood vessels from bone marrow progenitor cells.

doi: 10.1016/j.cardiores.2007.04.018

Figure Lengend Snippet: Fig. 1. BM-SMPC express biochemical markers of smooth muscle cells. (A) RT-PCR showed that BM-SMPC expressed smooth muscle cell markers as indicated. (B) Western blot for smooth muscle α-actin and calponin; beta-actin served as loading control. (C) Immunocytochemistry for smooth muscle α-actin and calponin. V-SMC from umbilical veins of near-term lambs were used as positive control. Representative results from three independent experiments are shown.

Article Snippet: Western blots were performed as described previously [33,34] using the following antibodies: mouse anti human smooth muscle α-actin (1:100 dilution; SeroTec) and mouse anti-human calponin (1:100 dilution; DakoCytomation) in TBS-Tween.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Immunocytochemistry, Positive Control

Journal: Cardiovascular research

Article Title: Functional tissue-engineered blood vessels from bone marrow progenitor cells.

doi: 10.1016/j.cardiores.2007.04.018

Figure Lengend Snippet: Fig. 2. BM-TEV displayed similar morphologic and biochemical characteristics as TEVs from V-SMC. BM-SMPC or V-SMC were embedded in fibrin hydrogels and cultured around 4-mm mandrel for 2 weeks to form cylindrical tubes. (A) Hematoxylin and eosin (H&E) staining showed that BM-SMPC were distributed uniformly in fibrin hydrogels (bar=100 μm). (B) Immunostaining of BM-TEV and TEV from V-SMC for smooth muscle α-actin and calponin (bar=100 μm).

Article Snippet: Western blots were performed as described previously [33,34] using the following antibodies: mouse anti human smooth muscle α-actin (1:100 dilution; SeroTec) and mouse anti-human calponin (1:100 dilution; DakoCytomation) in TBS-Tween.

Techniques: Cell Culture, Staining, Immunostaining