Journal: Cancer cell international

Article Title: RUNX2 interacts with BRG1 to target CD44 for promoting invasion and migration of colorectal cancer cells.

doi: 10.1186/s12935-020-01544-w

Figure Lengend Snippet: Fig. 3 RUNX2 interacts with BRG1 to promote CD44 transcription. a Both the siRNAs for BRG1 could effectively inhibit the expression of BRG1. b BRG1 knockdown decreased the expression of CD44. c BRG1 knockdown inhibited the sphere-forming ability of RKO cells. d Knockdown of BRG1 offset the ability of RUNX2 to promote the expression of CD44 in HT115 cells. e The analysis of RUNX2 binding to the CD44 promoter by CHIP assay in RKO cells. Genomic DNA was immunoprecipitated with anti-RUNX2 antibody, where IgG served as a negative control. The primer set specific to the RUNX2 binding site of CD44 promoter was used in real-time PCR amplification. Data are shown as means ± SE; *P<0.05, **P<0.01; NC negative control, si short interfering, RUNX2 overexpression of RUNX2

Article Snippet: The samples were then rinsed thrice with PBS, permeabilized with 0.25% Triton X-100 (9002931; AMRESCO, USA) in PBS for 10 min, and blocked with 5% normal goat serum (ZLI-9056; ZSGB-bio) in PBS for 60 min. After blocking, the cells were incubated with the corresponding primary antibodies RUNX2 (ab76956; Abcam, UK) and BRG1 (21634–1-AP; Proteintech, USA) overnight at 4 °C, followed by treatment with antimouse-Alexa Fluor 594 and anti-rabbit-Alexa Fluor 488 secondary antibody, respectively, for 1 h at the room temperature.

Techniques: Expressing, Knockdown, Binding Assay, Immunoprecipitation, Negative Control, Real-time Polymerase Chain Reaction, Amplification, Over Expression

Journal: Cancer cell international

Article Title: RUNX2 interacts with BRG1 to target CD44 for promoting invasion and migration of colorectal cancer cells.

doi: 10.1186/s12935-020-01544-w

Figure Lengend Snippet: Fig. 4 RUNX2 interacts with BRG1. a Confocal microscopy results showing that both RUNX2 and BRG1 were expressed in the nucleus of RKO and HT115 cells. b Co-IP experiments were performed with cell lysates of HEK293T expressing RUNX2-Myc and/or BRG1-GFP. The transfected constructs are indicated in the top panel. Anti-GFP (GFP-IP; left panel) or anti-Myc (Myc-IP; right panel) antibody was used for the immunoprecipitation assay. The antibodies used for WB are described on the left panel. c Co-IP experiments were performed on RKO cell lysates. The immunoprecipitation assay of RKO cell lysates with RUNX2, BRG1, or control (IgG) antibody. RUNX2 or BRG1 was detected by WB using the indicated antibody

Article Snippet: The samples were then rinsed thrice with PBS, permeabilized with 0.25% Triton X-100 (9002931; AMRESCO, USA) in PBS for 10 min, and blocked with 5% normal goat serum (ZLI-9056; ZSGB-bio) in PBS for 60 min. After blocking, the cells were incubated with the corresponding primary antibodies RUNX2 (ab76956; Abcam, UK) and BRG1 (21634–1-AP; Proteintech, USA) overnight at 4 °C, followed by treatment with antimouse-Alexa Fluor 594 and anti-rabbit-Alexa Fluor 488 secondary antibody, respectively, for 1 h at the room temperature.

Techniques: Confocal Microscopy, Co-Immunoprecipitation Assay, Expressing, Transfection, Construct, Immunoprecipitation, Control

Journal: Cancer cell international

Article Title: RUNX2 interacts with BRG1 to target CD44 for promoting invasion and migration of colorectal cancer cells.

doi: 10.1186/s12935-020-01544-w

Figure Lengend Snippet: Fig. 6 Model of the interaction and relationship among RUNX2, BRG1, CD44, and CRC. RUNX2 synergizes with BRG1 for promoting the CD44 expression as well as for enhancing the EMT process and the invasion and migration of CRC cells. Knockdown of BRG1 with specific si-RNA could markedly inhibit the RUNX2-mediated upregulation of the CD44 expression, suggesting that RUNX2 promotes the CD44 expression and EMT depending on the interaction between RUNX2 and BRG1 in CRC cells

Article Snippet: The samples were then rinsed thrice with PBS, permeabilized with 0.25% Triton X-100 (9002931; AMRESCO, USA) in PBS for 10 min, and blocked with 5% normal goat serum (ZLI-9056; ZSGB-bio) in PBS for 60 min. After blocking, the cells were incubated with the corresponding primary antibodies RUNX2 (ab76956; Abcam, UK) and BRG1 (21634–1-AP; Proteintech, USA) overnight at 4 °C, followed by treatment with antimouse-Alexa Fluor 594 and anti-rabbit-Alexa Fluor 488 secondary antibody, respectively, for 1 h at the room temperature.

Techniques: Expressing, Migration, Knockdown