Generating shRNA Knockdown Cell Lines

shRNA targeting sequences against IL-11 (XM_007998152.1) were designed using online design tools BLOCK-It RNAi Designer (Life Technologies), and three short target sequences with the best scores were selected. Targeting sequences are described in Additional file 1. The shRNA were cloned into the pLVX-shRNA1 vector (Takara, Dalian, China) containing EcoRI and BamHI sites. Lentiviral were produced in 293T cells following a standard liposome transfection with three packaging plasmids [24 (link)]. Then cells were incubated with the lentiviral particles (MOI 1) in the presence of 8 μg/mL Polybrene (Sigma-Aldrich). Forty-eight hours after infection, the cells were incubated with 6 μg/mL puromycin for selection. IL-11 target-specific silenced cell lines and the negative control (NC) scrambled vector-infected cells were named shRNA-IL-11 knock know (IL-11KD) and shRNA-negative control (shRNA-NC), respectively.