Journal: Cancers

Article Title: Lambda-Carrageenan Enhances the Effects of Radiation Therapy in Cancer Treatment by Suppressing Cancer Cell Invasion and Metastasis through Racgap1 Inhibition

doi: 10.3390/cancers11081192

Figure Lengend Snippet: Upregulation of RacGAP1 is involved in cancer cell survival and invasion after IR. ( A ) Gene expression was analyzed by cDNA microarray. RacGAP1 expression level is shown by the heat map. The values were normalized to the untreated group. ( B ) RacGAP1 protein expressions with different doses of IR were analyzed in MDA-MB-231, FaDu, PANC-1, and 4T1 cell lines. ( C ) MDA-MB-231 cells were transfected with siRNA duplexes targeting RacGAP1 (# 1 or # 2) or the control sequence, as indicated. Cell lysates were subjected to western blot two days after transfection. ( D – H ) MDA-MB-231 cells were transfected with siRNAs and incubated for two days, and then treated with 4 Gy IR. Each experiment was performed 24 h after IR treatment. Cell viability was quantified by cell counting ( D ). Apoptotic cells were measured by Annexin V-FITC staining and flow cytometry ( E ). ROS was measured by DCFDA ( F ). Cells were treated with staining solution and PI, and then the cell cycle was analyzed by flow cytometry. The percentage of polyploid cells in each group was normalized with control group ( G ). The invasive activity was measured by Matrigel chemoinvasion assay ( H ). Columns, mean ( n ≥ 3); bars, SE. *, p < 0.05; **, p < 0.01. Data shown in ( D – H ) were normalized to that of MDA-MB-231 cells transfected with siRNA control. For the continuous full length-image of Western blot signals, please refer to .

Article Snippet: Each slide was incubated in 2% blocking buffer (Roche, Basel, Switzerland) for 1 h, and then incubated with a RacGAP1 primary antibody (Proteintech) overnight.

Techniques: Gene Expression, Microarray, Expressing, Transfection, Control, Sequencing, Western Blot, Incubation, Cell Counting, Staining, Flow Cytometry, Activity Assay

Journal: Cancers

Article Title: Lambda-Carrageenan Enhances the Effects of Radiation Therapy in Cancer Treatment by Suppressing Cancer Cell Invasion and Metastasis through Racgap1 Inhibition

doi: 10.3390/cancers11081192

Figure Lengend Snippet: RacGAP1 expression is suppressed by CGN in MDA-MB-231 cells. ( A ) Protein expression of RacGAP1 was analyzed after treatment with different concentrations of CGN in MDA-MB-231 cells. Total cell lysates were subjected to western blot. ( B ) RacGAP1 protein expression was analyzed using cell lysates after IR and/or CGN treatment in MDA-MB-231 cells. ( C ) Immunofluorescence images show RacGAP1 (green), α-tubulin (red) and nuclei (blue). Bar, 25 μm. ( D ) RacGAP1-mVenus expression was induced by doxycycline in MDA-MB-231 cells. RacGAP1 expression was analyzed by western blot using anti-RacGAP1 antibody. Dox, doxycycline. ( E ) Cell number was quantified by cell counting. Columns, mean ( n = 3); bars, SE. *, p < 0.05; **, p < 0.01. For the continuous full length-image of Western blot signals, please refer to .

Article Snippet: Each slide was incubated in 2% blocking buffer (Roche, Basel, Switzerland) for 1 h, and then incubated with a RacGAP1 primary antibody (Proteintech) overnight.

Techniques: Expressing, Western Blot, Immunofluorescence, Cell Counting

Journal: Cancers

Article Title: Lambda-Carrageenan Enhances the Effects of Radiation Therapy in Cancer Treatment by Suppressing Cancer Cell Invasion and Metastasis through Racgap1 Inhibition

doi: 10.3390/cancers11081192

Figure Lengend Snippet: Radiation followed by CGN treatment decreases tumor size and metastasis in a 4T1 mouse xenograft model. ( A ) Treatment schedule of each group. Mouse 4T1 cells were injected into Balb/c mice ( n = 6–7 in each group). ( B ) 4T1 tumor sizes were measured and normalized to size at day 12 in each group. ( C ) Relative tumor size for each group was measured on Day 25. Scatter plot; mean ± SE. *, p < 0.05. ( D ) Representative H&E images of lung sections. Bar, 500 µm. Arrow, metastatic lung nodules. The number of metastatic lung nodules was counted (right panel). Scatter plot; mean ± SE. *, p < 0.05. ( E ) Sections from 4T1 tumors were subjected to IHC staining with antibodies against RacGAP1. Bar, 100 µm. RacGAP1 expression level was determined by scoring, as described in the methods (right panel). Scatter plot; bars, mean ( n = 5). *, p < 0.05.

Article Snippet: Each slide was incubated in 2% blocking buffer (Roche, Basel, Switzerland) for 1 h, and then incubated with a RacGAP1 primary antibody (Proteintech) overnight.

Techniques: Injection, Immunohistochemistry, Expressing