Pgfp 5 rs

Reagents involved in cell cultures were purchased from Thermo Fisher Scientific. Pharmacological inhibitors including rapamycin, rapalogs and other chemical inhibitors were purchased from LC Laboratories. Stable cell line CER was used to obtain the recombinant reelin in the conditioned medium as described [32] (link), and the corresponding mock medium was obtained from the parental 293 EBNA cell line as described [31] (link). Maf1 specific 29mer shRNA constructs were expressed from retroviral GFP vector (pGFP-V-RS) (OriGene). Maf1 shRNA and scramble shRNA were expressed from AAV-PHP.eB vectors (PackGene Biotech). Vector overexpressing Myc-tagged Maf1 was constructed from pCMV6-AN-His-Myc vector (OriGene). Raptor shRNA was expressed from pLKO.1 vector (AddGene). Antibodies and other experimental reagents used in this study can be found in Table S3.

Plasmids encoding human Notch1 shRNA, empty vector (pGFP‐V‐RS), and TurboFectin 8.0 transfection reagent, were purchased from OriGene Technologies (Rockville, MD, USA). PC‐9 and H1975 cells were divided equally into two groups: shNotch1 (transfected with Notch1 shRNA) and shControl (transfected with empty vector) cells. The day prior to transfection, cells were seeded at a density of 1 × 10⁵/well onto six‐well plates. Then, the cells were transfected with 2 μg Notch1 shRNA plasmid or blank vector in serum‐free Opti‐MEM I (Thermo Fisher Scientific) using 6 μl TurboFectin 8.0. After 24 h, the transfected cells were diluted 1:10 in 10‐cm dishes and the culture medium was replaced with complete medium containing puromycin (Sigma‐Aldrich). Stably positive clones were obtained after screening with puromycin.