artemisinin treated cells  (Santa Cruz Biotechnology)

Journal: BMC cancer

Article Title: Transcriptome analysis of genes associated with breast cancer cell motility in response to Artemisinin treatment.

doi: 10.1186/s12885-017-3863-7

Figure Lengend Snippet: Fig. 1 Artemisinin inhibits growth and colony forming ability of estrogen receptor positive breast cancer cells. (A) Viability assay in MCF10A, MCF-7, T47D and MDA-MB-231 breast cancer cells showing the effect of artemisinin treatment in a dose and time dependent manner where artemisinin concentration is indicated in X axis and percentage viability compared to control is indicated on the Y axis. The mean + SEM for three independent experiments was calculated. Statistically significant difference was found between the absorbance of control and artemisinin treated samples ***p (<0.001), **p (<0.0078) and ns p (>0.05). B (I) Representative image of colony forming assay of artemisinin treated MCF10A, MCF-7, T47D and MDA-MB-231 breast cancer cells. (II) Graph represents mean + SEM of control, and treated samples in three separate experiments performed in triplicate, *p(<0.05), ***p (<0.001)

Article Snippet: The 10 μM artemisinin treated cells were washed with ice-cold 1X PBS, fixed with methanol:acetone (1:1) and kept at -20 °C for 30 min-1 h. The cells were then blocked with blocking buffer [0.1% (w/v) bovine serum albumin, 0.3% (v/v) TritonTM X-100 in 1X PBS] for 2 h and then incubated with primary antibodies [Cytochrome c antibody (1:500, Santa Cruz), β catenin (1:5000)] overnight at 4 °C.

Techniques: Viability Assay, Concentration Assay, Control

Journal: BMC cancer

Article Title: Transcriptome analysis of genes associated with breast cancer cell motility in response to Artemisinin treatment.

doi: 10.1186/s12885-017-3863-7

Figure Lengend Snippet: Fig. 2 Artemisinin exhibits anti-migratory, anti-invasion and apoptosis inducing property in breast cancer cells. A (I) Picture represent relative cell migration in both control and treated MCF-7 cells at different time intervals. (II) Graph represents the quantification of the decrease in the area as wound healing progresses at the observed time points. Significant differences were observed between control and treated cells at different time points p (<0.0001). B (I) Image depicts the cell migration in control and artemisinin treated MCF7 cells as observed in transwell migration assay. (II) Graph depicts the average number of migrated cells. C (I) Diagram represents relative invasion in control and artemisinin treated aggressive breast cancer cells. (II) Relative invasion in depicted in the graph. D (I) Dot plot representing PE Annexin V positive, 7AAD negative MCF-7 cells after 24 h of treatment with 1 μM artemisinin, control (DMSO < 0.01%)μ and plumbagin (5 μM) as positive control. The lower left quadrants of each panels show the viable cells and 7-AAD negative, lower right quadrants represent the early apoptotic cells (PE Annexin V positive and 7-AAD negative). (II) Graph represents the percentage of early apoptotic cells in control and artemisinin treated MCF-7 cells computed from three biologically different set of experiments. Significant differences were observed between control and treated cells, *p < 0.05

Article Snippet: The 10 μM artemisinin treated cells were washed with ice-cold 1X PBS, fixed with methanol:acetone (1:1) and kept at -20 °C for 30 min-1 h. The cells were then blocked with blocking buffer [0.1% (w/v) bovine serum albumin, 0.3% (v/v) TritonTM X-100 in 1X PBS] for 2 h and then incubated with primary antibodies [Cytochrome c antibody (1:500, Santa Cruz), β catenin (1:5000)] overnight at 4 °C.

Techniques: Migration, Control, Transwell Migration Assay, Positive Control

Journal: BMC cancer

Article Title: Transcriptome analysis of genes associated with breast cancer cell motility in response to Artemisinin treatment.

doi: 10.1186/s12885-017-3863-7

Figure Lengend Snippet: Fig. 3 Artemisinin alters the expression of genes associated with growth promoting activities. a Heat map showing the fold change expression of genes under study. b and c qRT PCR and western blot assay respectively showing the expression of genes associated with mammary gland development upon artemisinin treatment. d and e Respective RNA and protein expression of cell proliferation associated genes in control and artemisinin treated MCF7 cells. f and g Bar diagram and immunoblot respectively showing the expression level of proteins involved in migration, invasion and apoptosis in artemisinin treated and control cells

Article Snippet: The 10 μM artemisinin treated cells were washed with ice-cold 1X PBS, fixed with methanol:acetone (1:1) and kept at -20 °C for 30 min-1 h. The cells were then blocked with blocking buffer [0.1% (w/v) bovine serum albumin, 0.3% (v/v) TritonTM X-100 in 1X PBS] for 2 h and then incubated with primary antibodies [Cytochrome c antibody (1:500, Santa Cruz), β catenin (1:5000)] overnight at 4 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Migration

Journal: BMC cancer

Article Title: Transcriptome analysis of genes associated with breast cancer cell motility in response to Artemisinin treatment.

doi: 10.1186/s12885-017-3863-7

Figure Lengend Snippet: Fig. 4 Cytoplasmic localization of Beta-catenin increases upon artemisinin treatment in MCF-7 breast cancer cells. a Immunofluorescence assay shows the expression and localization of beta-catenin upon artemisinin treatment. b Immunoblot against beta-catenin shows the increased cytoplasmic beta-catenin protein expression upon artemisinin treatment. α-tubulin and histone H3 used as loading control

Article Snippet: The 10 μM artemisinin treated cells were washed with ice-cold 1X PBS, fixed with methanol:acetone (1:1) and kept at -20 °C for 30 min-1 h. The cells were then blocked with blocking buffer [0.1% (w/v) bovine serum albumin, 0.3% (v/v) TritonTM X-100 in 1X PBS] for 2 h and then incubated with primary antibodies [Cytochrome c antibody (1:500, Santa Cruz), β catenin (1:5000)] overnight at 4 °C.

Techniques: Immunofluorescence, Expressing, Western Blot, Control

Journal: BMC cancer

Article Title: Transcriptome analysis of genes associated with breast cancer cell motility in response to Artemisinin treatment.

doi: 10.1186/s12885-017-3863-7

Figure Lengend Snippet: Fig. 5 Artemisinin induced apoptosis in MCF-7 cells is also through increased Cytochrome c release and Caspase 9 cleavage. a Confocal images of cytochrome c release. Cells stained with mitotracker DiOC6 [91], Cytochrome c (red), merged image shows Cytochrome c release (yellow). b Immunoblot against cytochrome c, and caspase 9 showing increased cleaved caspase 9

Article Snippet: The 10 μM artemisinin treated cells were washed with ice-cold 1X PBS, fixed with methanol:acetone (1:1) and kept at -20 °C for 30 min-1 h. The cells were then blocked with blocking buffer [0.1% (w/v) bovine serum albumin, 0.3% (v/v) TritonTM X-100 in 1X PBS] for 2 h and then incubated with primary antibodies [Cytochrome c antibody (1:500, Santa Cruz), β catenin (1:5000)] overnight at 4 °C.

Techniques: Staining, Western Blot

Journal: BMC cancer

Article Title: Transcriptome analysis of genes associated with breast cancer cell motility in response to Artemisinin treatment.

doi: 10.1186/s12885-017-3863-7

Figure Lengend Snippet: Fig. 6 Artemisinin inhibits HDACs. A (I) Western blot assay with protein extracted from MCF-7, T47D and MDA-MB-231 cells treated with artemisinin. Immunoblot was developed using primary antibody HDAC 1, 2, 3 and 6. (II) Densitometry analysis of the protein levels of HDACs as observed in the western blot

Article Snippet: The 10 μM artemisinin treated cells were washed with ice-cold 1X PBS, fixed with methanol:acetone (1:1) and kept at -20 °C for 30 min-1 h. The cells were then blocked with blocking buffer [0.1% (w/v) bovine serum albumin, 0.3% (v/v) TritonTM X-100 in 1X PBS] for 2 h and then incubated with primary antibodies [Cytochrome c antibody (1:500, Santa Cruz), β catenin (1:5000)] overnight at 4 °C.

Techniques: Western Blot