Journal: PLOS ONE

Article Title: Shenqi Fuzheng injection reverses M2 macrophage-mediated cisplatin resistance through the PI3K pathway in breast cancer

doi: 10.1371/journal.pone.0279752

Figure Lengend Snippet: Tumour lysates were analysed for the expressions of p-PI3K, PI3K, P-gp, and ABCG2 with the respective antibodies. The density ratio of detected proteins to GAPDH is shown as relative expression. Values are shown as the means ± SD from three independent experiments. *p < 0.05 as compared to Cisplatin alone.

Article Snippet: Monoclonal antibodies against CD206, CD86, PI3K, P-gp, ABCG2, Bcl-2, Bax, and GAPDH were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Expressing

Journal: PLOS ONE

Article Title: Shenqi Fuzheng injection reverses M2 macrophage-mediated cisplatin resistance through the PI3K pathway in breast cancer

doi: 10.1371/journal.pone.0279752

Figure Lengend Snippet: Cell lysates were analysed for the expressions of P-gp, ABCG2, Bcl-2, and Bax with the respective antibodies. The density ratio of detected proteins to GAPDH is shown as relative expression. Values are shown as the means ± SD from three independent experiments. *p < 0.05 as compared to the Control. #p < 0.05 as compared to IGF-1 alone.

Article Snippet: Monoclonal antibodies against CD206, CD86, PI3K, P-gp, ABCG2, Bcl-2, Bax, and GAPDH were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Expressing

Journal: Gastroenterology Research and Practice

Article Title: Expression of Potential Cancer Stem Cell Marker ABCG2 is Associated with Malignant Behaviors of Hepatocellular Carcinoma

doi: 10.1155/2013/782581

Figure Lengend Snippet: Expression of ABCG2 in human HCC tissues. (a) ABCG2 was expressed in placenta tissues (positive control). (b) Primary antibody omitted placenta tissue slide was set as negative control. (c) Positive ABCG2 expression in HCC tissue. (d) Negative ABCG2 expression in HCC tissue.

Article Snippet: Sections were incubated with rabbit anti-ABCG2 polyclonal antibody (1 : 200, 4477S, Cell Signaling Technology, Danvers, MA) and rabbit anti-Ki67 monoclonal antibody (1 : 900, ab16667, Abcam, Cambridge, MA) at 4°C overnight.

Techniques: Expressing, Positive Control, Negative Control

Journal: Gastroenterology Research and Practice

Article Title: Expression of Potential Cancer Stem Cell Marker ABCG2 is Associated with Malignant Behaviors of Hepatocellular Carcinoma

doi: 10.1155/2013/782581

Figure Lengend Snippet: ABCG2 expression and characteristics of patients.

Article Snippet: Sections were incubated with rabbit anti-ABCG2 polyclonal antibody (1 : 200, 4477S, Cell Signaling Technology, Danvers, MA) and rabbit anti-Ki67 monoclonal antibody (1 : 900, ab16667, Abcam, Cambridge, MA) at 4°C overnight.

Techniques: Expressing

Journal: Gastroenterology Research and Practice

Article Title: Expression of Potential Cancer Stem Cell Marker ABCG2 is Associated with Malignant Behaviors of Hepatocellular Carcinoma

doi: 10.1155/2013/782581

Figure Lengend Snippet: ABCG2 was expressed in a minor population of SMMC-7721 cells. (a) The positive rate of ABCG2 in SMMC-7721 cells determined by flow cytometry was about 8.8% before cell sorting. P2 gate represents ABCG2 positive cells. Mouse IgG2b was used as isotype control. (b) Positive rate of ABCG2 in sorted ABCG2+/ABCG2− cells after 3 passages of culture. (c) Detection of ABCG2 protein level in freshly sorted cells by western blot. β -Actin was used as loading control. The optical density of specific bands was quantified and normalized to β -Actin. ** P < 0.01.

Article Snippet: Sections were incubated with rabbit anti-ABCG2 polyclonal antibody (1 : 200, 4477S, Cell Signaling Technology, Danvers, MA) and rabbit anti-Ki67 monoclonal antibody (1 : 900, ab16667, Abcam, Cambridge, MA) at 4°C overnight.

Techniques: Flow Cytometry, FACS, Western Blot

Journal: Gastroenterology Research and Practice

Article Title: Expression of Potential Cancer Stem Cell Marker ABCG2 is Associated with Malignant Behaviors of Hepatocellular Carcinoma

doi: 10.1155/2013/782581

Figure Lengend Snippet: Difference of tumorigenic ability between ABCG2+ and ABCG2− cells.

Article Snippet: Sections were incubated with rabbit anti-ABCG2 polyclonal antibody (1 : 200, 4477S, Cell Signaling Technology, Danvers, MA) and rabbit anti-Ki67 monoclonal antibody (1 : 900, ab16667, Abcam, Cambridge, MA) at 4°C overnight.

Techniques:

Journal: Gastroenterology Research and Practice

Article Title: Expression of Potential Cancer Stem Cell Marker ABCG2 is Associated with Malignant Behaviors of Hepatocellular Carcinoma

doi: 10.1155/2013/782581

Figure Lengend Snippet: Expression level of ABCG2 correlated with cell proliferation and chemoresistance. (a), (b) Freshly sorted ABCG2+ cells were transfected with ABCG2-specific siRNA. ABCG2− cells were transfected with ABCG2-overexpression plasmid. The proliferation of cells after transfection was compared with untransfected cells and negative control (scrambled siRNA or empty plasmid) using MTT assay after indicated time. (c), (d) The expression level of ABCG2 was manipulated as described above. Survival rates after exposure to indicated dose of doxorubicin for 24 h were determined by MTT assay. (e), (f) IC50 value to doxorubicin was calculated for different groups. ** P < 0.01.

Article Snippet: Sections were incubated with rabbit anti-ABCG2 polyclonal antibody (1 : 200, 4477S, Cell Signaling Technology, Danvers, MA) and rabbit anti-Ki67 monoclonal antibody (1 : 900, ab16667, Abcam, Cambridge, MA) at 4°C overnight.

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Negative Control, MTT Assay

Journal: Gastroenterology Research and Practice

Article Title: Expression of Potential Cancer Stem Cell Marker ABCG2 is Associated with Malignant Behaviors of Hepatocellular Carcinoma

doi: 10.1155/2013/782581

Figure Lengend Snippet: RNA interference and plasmid-mediated overexpression of ABCG2 in SMMC-7721 cells. (a) RT-PCR analysis of ABCG2 mRNA in SMCC-7721 cells after transfection with siRNA or plasmid for 48 h. Normal SMMC-7721 cells were used as blank control. Scrambled siRNA was set as negative control. For overexpression, empty plasmid was transfected as negative control. (b), (c) Western blot analysis confirmed the efficiency of downregulation of ABCG2 by siRNA and upregulation by overexpression, respectively. β -Actin served as loading control. The optical density was quantified and normalized to β -Actin. ** P < 0.01, *** P < 0.001.

Article Snippet: Sections were incubated with rabbit anti-ABCG2 polyclonal antibody (1 : 200, 4477S, Cell Signaling Technology, Danvers, MA) and rabbit anti-Ki67 monoclonal antibody (1 : 900, ab16667, Abcam, Cambridge, MA) at 4°C overnight.

Techniques: Plasmid Preparation, Over Expression, Reverse Transcription Polymerase Chain Reaction, Transfection, Negative Control, Western Blot

Journal: Gastroenterology Research and Practice

Article Title: Expression of Potential Cancer Stem Cell Marker ABCG2 is Associated with Malignant Behaviors of Hepatocellular Carcinoma

doi: 10.1155/2013/782581

Figure Lengend Snippet: Downregulation or upregulation of ABCG2 had effect on migration and invasion potential in HCC cells. (a), (b) Wound healing assay evaluated the potential of migration of HCC cells. Untreated cells served as normal control. ABCG2+ cells were transfected with ABCG2 siRNA while ABCG2− cells were transfected with ABCG2-expressing plasmid. Scrambled siRNA and empty plasmid were used as negative control. The pictures above represent the wound right after tip scratch. Pictures below showed wound-healing status after 48 h. (c), (d) Results of transwell invasion assay. After 48 h of invasion, cells invaded to the lower chamber were stained and photographed. Invaded cells were counted in three random visions and the average numbers were presented. *** P < 0.001.

Article Snippet: Sections were incubated with rabbit anti-ABCG2 polyclonal antibody (1 : 200, 4477S, Cell Signaling Technology, Danvers, MA) and rabbit anti-Ki67 monoclonal antibody (1 : 900, ab16667, Abcam, Cambridge, MA) at 4°C overnight.

Techniques: Migration, Wound Healing Assay, Transfection, Expressing, Plasmid Preparation, Negative Control, Transwell Invasion Assay, Staining