Fig. 2.

Suppression of tumor cell growth after shRNA-mediated gene knockdown of TdIF1 in A549 cells. a Knockdown of TdIF1 in A549 lung cancer cells via TdIF1-shRNA lentiviral vector. The lentiviral vector is a hU6-MCS-CMV-EGFP construct. The RNA sequence (TTCTCCGAACGTGTCACGT) was synthesized and ligated into the lentiviral vector. A549 cells were cultured and transfected with the TdIF1 shRNA lentiviral vector (shTdIF1) or a control nonspecific shRNA vector (shCtrl) at an MOI of 10 for 24 h. TdIF1 mRNA was detected by qPCR, and TdIF1 protein expression was detected by western blot. b In vitro suppression of cell proliferation by gene silencing of TdIF1. A549 cells were transfected with the lentiviral vector shTdIF1 or the control vector shCtrl at an MOI of 10 for 24 h. Live cells were stained with a green dye and detected by a Celigo image cytometer for 5 days. Cell images, cell number counts and cell proliferation fold changes are presented. c Decreased colony formation of A549 cells after gene knockdown of TdIF1. A549 cells were transfected with the lentiviral vector shTdIF1 or the control vector shCtrl at an MOI of 1:100 for 24 h. Approximately 1000 cells were plated in 6-well plates and cultured for 11 days. The cell clones were determined by staining with crystal violet. The crystal violet-stained colonies were imaged and counted (presented as a percentage) using a microscope. Error bars represent the standard deviation of three experiments (*P < 0.05; **P < 0.01)