cst cat 51917  (Cell Signaling Technology Inc)

Journal: Molecular carcinogenesis

Article Title: Oncogenic fusion of CD63-BCAR4 contributes cancer stem cell-like properties via ALDH1 activity.

doi: 10.1002/mc.23808

Figure Lengend Snippet: FIGURE 2 Cancer stemness markers are upregulated in CD63‐BCAR4 overexpressed cells. (A) Altered expression of cancer stem cell (CSCs) markers analyzed by RNA microarray in overexpressed cell lines of BCAR4 fusion. Genes showing increased expression are shown in red with a gradient based on fold difference. (B) Relative expression of ALDH1A1, CD133, and CD44 in mRNA level analyzed by qRT‐PCR. (C) Flow cytometry detection of CD44‐, CD133‐, and ALDH1A1‐expressing cells. The experiment was conducted three times, and the results were presented as the mean value ± SEM (standard error of the mean). (D) Western blot analysis of CSC markers in each cell line and after siRNA transfection. Representative images from the replicated experiments are presented, and statistical significances are marked with (*p < 0.05), (**p < 0.005), (***p < 0.0005), and (****p < 0.00005). BCAR4, breast cancer antiestrogen resistance 4; qRT‐PCR, quantitative real‐time polymerase chain reaction. [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Next, the cells were incubated with the appropriate primary antibody (CD44, Cat#77862, Cell Signaling Technology [CST]; CD133, Cat#51917, CST; ALDH1A1, Cat#60171, Proteintech; OCT‐3/4, Cat#11263, Proteintech) for overnight at 4°C.

Techniques: Expressing, Microarray, Quantitative RT-PCR, Flow Cytometry, Western Blot, Transfection, Real-time Polymerase Chain Reaction

Journal: Molecular carcinogenesis

Article Title: Oncogenic fusion of CD63-BCAR4 contributes cancer stem cell-like properties via ALDH1 activity.

doi: 10.1002/mc.23808

Figure Lengend Snippet: FIGURE 3 ALDH1A1 is upregulated in the spheres of CD63‐BCAR4 expressed cells. (A) ALDH activity was measured by colorimetric assay in each stable cell line. (B) Protein expressions of stem cell markers in 2D cultured cells and spheres. (C) Immunofluorescence staining of CD44, CD133, and ALDH1A1 in the spheres after siRNA transfection. DAPI was used to stain the nucleus, and images were from the representative data. (D) The graph shows the mean values of the intensity of each protein from the replicated experiments. Statistical significances are marked with (*p < 0.05), (**p < 0.005), and (***p < 0.0005). BCAR4, breast cancer antiestrogen resistance 4. [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Next, the cells were incubated with the appropriate primary antibody (CD44, Cat#77862, Cell Signaling Technology [CST]; CD133, Cat#51917, CST; ALDH1A1, Cat#60171, Proteintech; OCT‐3/4, Cat#11263, Proteintech) for overnight at 4°C.

Techniques: Activity Assay, Colorimetric Assay, Stable Transfection, Cell Culture, Immunofluorescence, Staining, Transfection