Article Title: Slit2 inhibits glioma cell invasion in the brain by suppression of Cdc42 activity
Figure Lengend Snippet: Expression of Slit2 and Robo1 in primary glioma specimens and glioma cell lines. (A) Slit2 and Robo1 are expressed at lower levels in primary human glioma specimens and invasive SNB19 and U373MG glioma cells compared with normal human brain tissue (NB) and normal human astrocytes (NHA). Synthesized first-strand cDNA from total RNA of NB, primary glioma specimens (J4, J5, glioblastoma multiforme, WHO grade IV; J15 and J16, anaplastic oligodendroglioma, grade III), NHA, and SNB19 and U373MG glioma cells was used for PCR reactions. The length of synthesized cDNA fragments is 514 bp for Robo1 and 387 bp for Slit2, respectively. β-Actin was used as internal control. (B) Ectopic expression of Slit2 in SNB19 and U373MG glioma cells. Cell lysates (top two panels) or conditioned media (CM, bottom panel) from SNB19, SNB19 Slit2-expressing, U373MG, or U373MG Slit2-expressing cells were analyzed by immunoblot using an anti-c-Myc antibody. Lanes 2, 3, and 4 are SNB19 Slit2-expressing cell clones; lanes 7, 56, and 38 are U373MG Slit2-expressing cell clones. β-Actin was used as a loading control. Results shown in A and B are representative of three independent experiments.
Article Snippet: The following antibodies were used in our studies: goat anti-β-actin (I-19, SC-1616) and mouse anti-c-Myc (9E10, SC-40 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-β-catenin (610153; BD Biosciences, San Diego, CA, USA); anti-N-cadherin (3B9, 33-3900; Invitrogen Life Science Zymed, Carlsbad, CA, USA); antiphospho-tyrosine (4G10, 05-321, Upstate Biotechnology, Lake Placid, NY, USA); and mouse anti-N-Cadherin (GC-4, C3865; Sigma Chemicals, St. Louis, MO, USA).
Techniques: Expressing, Synthesized, Polymerase Chain Reaction, Clone Assay