Mission shrna targeting set

SALL3 KD cells were generating by infecting 253G1 and 201B7 cells with lentiviral particles expressing SALL3-targeted shRNAs. Briefly, Lenti-X 293T cells were transfected with individual clones from a Sigma MISSION shRNA targeting set (No. 1: TRCN0000019754, No. 2: TRCN0000417790) or control shRNA plasmid along with a MISSION Lentivirus Packaging Mix (Sigma-Aldrich) according to the manufacturer’s instructions. Media containing viruses were collected 48 h after transfection, and cells were transduced with the viruses in the presence of 8 µg mL⁻¹ polybrene (Sigma-Aldrich) for 24 h. The cells were then subjected to selection by 2 µg mL⁻¹ puromycin (Gibco) for 48 h. The mRNA microarray data of the control and SALL3 shRNA cells were obtained as described above.

ARPE-19 TNNT1-knockdown cells were generating by transducing ARPE-19 with lentivirus particles that expressed a TNNT1-targeting set of shRNA. Briefly, Lenti-X 293 T cells were transfected with individual clones from Sigma MISSION shRNA targeting set (TRCN0000118897) or the control shRNA plasmid along with a MISSION lentivirus packaging Mix (Sigma-Aldrich) according to the manufacture’s procedures. Media containing viruses were collected 48 hours after transfection. ARPE-19 cells were transduced with the viruses in the presence of 8 µg/ml polybrene (Sigma-Aldrich) for 24 hours, and then subjected to selection with 2 µg/ml puromycin for 48 hours.