Journal: Cell Reports

Article Title: An integrated model for Gpr124 function in Wnt7a/b signaling among vertebrates

doi: 10.1016/j.celrep.2022.110902

Figure Lengend Snippet:

Article Snippet: pRK5- mFZ1 , ( ) , Addgene #42253.

Techniques: Recombinant, Transfection, Protease Inhibitor, Western Blot, Labeling, Luciferase, Reporter Assay, Clone Assay, Software, CRISPR, Expressing

Journal: PLoS ONE

Article Title: Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

doi: 10.1371/journal.pone.0198508

Figure Lengend Snippet: (A) Survival curve of glioblastoma cells after irradiation. The cells were irradiated with up to 8 Gy. (B) Quantification of Robo1-, Slit2-mRNA expression by qPCR. Values represent means ± standard deviation and are relative to the expression of the housekeeping gene GAPDH (100%). ****p<0.0001, **p<0.01 (Student t-test). (C) Motility of non-irradiated U-373 MG and U-87 MG cells. Cell motility was measured by the parameters velocity, Euclidean distance, and accumulated total distance. Data shown are means ± standard error. ***p<0.001 (Mann-Whitney U test).

Article Snippet: U-373 MG and U-87 MG cells were transfected with 80 pmol Robo1 small interfering RNA (siRNA; Cat.No. sc-42252, Santa Cruz Biotechnology, USA) or mock siRNA (control sample; Cat.No. sc-37007) using Lipofectamine LTX following the kit’s instructions.

Techniques: Irradiation, Expressing, Standard Deviation, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

doi: 10.1371/journal.pone.0198508

Figure Lengend Snippet: Robo1 (A) and Slit2 (B) mRNA in U-373 MG cells quantified by qPCR 24h after photon irradiation. Relative expression levels were calculated using the ΔΔCt method (reference gene: GAPDH) and are shown relative to those in the non-irradiated controls (0 Gy, 100%). Data represent means ± standard deviation. **p<0.01, *p<0.05 (Student t-test).

Article Snippet: U-373 MG and U-87 MG cells were transfected with 80 pmol Robo1 small interfering RNA (siRNA; Cat.No. sc-42252, Santa Cruz Biotechnology, USA) or mock siRNA (control sample; Cat.No. sc-37007) using Lipofectamine LTX following the kit’s instructions.

Techniques: Irradiation, Expressing, Standard Deviation

Journal: PLoS ONE

Article Title: Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

doi: 10.1371/journal.pone.0198508

Figure Lengend Snippet: (A) Quantification of Robo1 and Slit2 mRNA expression in Robo1 and Slit2 overexpressing clones of the cell line U-373 MG. Relative expression levels were calculated using the ΔΔCt method (reference gene: GAPDH) and are shown relative to control (100%). Ctrl, U-373 MG-Control (control cells transfected with the empty vector). (C) Quantification of the adhesion of U-373 MG cells overexpressing Slit2 or Robo1 compared to that of control cells. Cells were treated either with trypsin/EDTA or EDTA alone. The adhesion index represents the percentage of adherent cells after the detached cells were removed. (B and D) Effect of irradiation and/or overexpression of Slit2 and Robo1 on the motility of U-373 MG cells. The motility of non-irradiated U-373 MG-Control, U-373 MG-Robo1, and U-373 MG-Slit2 cells (B, 0 Gy in D) or after irradiation with doses of 0.5 Gy, 2 Gy, and 8 Gy was compared. The cells were observed for 24h using time-lapse video microscopy and the motility parameters velocity, accumulated distance, and Euclidean distance were determined. Data represent means ± standard errors from at least four separate experiments. ****p<0.0001, ***p<0.001, **p<0.01, *p<0.05 (B and D, Mann-Whitney U test; C, ANOVA/Tukey).

Article Snippet: U-373 MG and U-87 MG cells were transfected with 80 pmol Robo1 small interfering RNA (siRNA; Cat.No. sc-42252, Santa Cruz Biotechnology, USA) or mock siRNA (control sample; Cat.No. sc-37007) using Lipofectamine LTX following the kit’s instructions.

Techniques: Expressing, Clone Assay, Transfection, Plasmid Preparation, Irradiation, Over Expression, Microscopy, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

doi: 10.1371/journal.pone.0198508

Figure Lengend Snippet: (A) Western blot assay demonstrating the downregulation of Robo1. WT, untreated control cells; siMock, siRobo1, cells transfected with control or Robo1-specific siRNAs. (B) The motility parameters velocity, accumulated distance, and Euclidean distance were determined in non-irradiated (0 Gy), Robo1-knockdown, and control U-373 MG cells and in Robo1-knockdown cells irradiated with a dose of 2 Gy. Data represent means ± standard errors. **p<0.01, *p<0.05 (Mann-Whitney U test).

Article Snippet: U-373 MG and U-87 MG cells were transfected with 80 pmol Robo1 small interfering RNA (siRNA; Cat.No. sc-42252, Santa Cruz Biotechnology, USA) or mock siRNA (control sample; Cat.No. sc-37007) using Lipofectamine LTX following the kit’s instructions.

Techniques: Western Blot, Transfection, Irradiation, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

doi: 10.1371/journal.pone.0198508

Figure Lengend Snippet: (A) Quantification of Robo1 mRNA expression in U-87 MG-Control cells (Ctrl) and U-87 MG-Robo1 cells (Robo1). Relative expression levels were calculated using the ΔΔCt method (reference gene: GAPDH) and are shown relative to control (Ctrl, 100%). (B) Motility of U-87 MG-Control and U-87 MG-Robo1 cells 24h after photon irradiation with doses ranging from 0 Gy to 8 Gy. Motility of the Robo1-overexpressing cells (solid line) is lower than that of wild-type cells (dashed line). (C) siRNA-knockdown of Robo1 in U-87 MG cells as demonstrated by Western blot analysis. (D) The motility parameters velocity, accumulated distance, and Euclidean distance were determined in non-irradiated (0 Gy) Robo1-knockdown and control U-87 MG cells and in Robo1-knockdown cells irradiated with a dose of 2 Gy. Data (B and D) represent means ± standard errors. ***p<0.001 (Mann-Whitney U test).

Article Snippet: U-373 MG and U-87 MG cells were transfected with 80 pmol Robo1 small interfering RNA (siRNA; Cat.No. sc-42252, Santa Cruz Biotechnology, USA) or mock siRNA (control sample; Cat.No. sc-37007) using Lipofectamine LTX following the kit’s instructions.

Techniques: Expressing, Irradiation, Western Blot, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Robo1 and vimentin regulate radiation-induced motility of human glioblastoma cells

doi: 10.1371/journal.pone.0198508

Figure Lengend Snippet: The expression of vimentin (A, E-G), fascin (B, H-J), focal adhesion kinase (FAK) (C, K-M), and phosphorylated focal adhesion kinase (pFAK Y925 ) (D, N-P) was quantified in U-373 MG wild-type cells and clones overexpressing Slit2 or Robo1 and the effect of photon irradiation on their expression was assessed. Proteins were isolated 24h after irradiation. WT, U-373 MG wild-type cells; Ctrl, U-373 MG control cells (transfected with empty vector); Robo1, U-373 MG-Robo1 (stable clone overexpressing Robo1); Slit2, U-373 MG-Slit2 (stable clone overexpressing Slit2).

Article Snippet: U-373 MG and U-87 MG cells were transfected with 80 pmol Robo1 small interfering RNA (siRNA; Cat.No. sc-42252, Santa Cruz Biotechnology, USA) or mock siRNA (control sample; Cat.No. sc-37007) using Lipofectamine LTX following the kit’s instructions.

Techniques: Expressing, Clone Assay, Irradiation, Isolation, Transfection, Plasmid Preparation, Stable Transfection