Fig. 6

FSTL1 links YAP and TRAM2 to EMT. a ChIP-qPCR was used to quantify YAP binding to the indicated regions. NC1 and PC (promoter of ANKRD1) are negative and positive control regions, respectively. Relative YAP-binding enrichment was calculated and normalized to NC1 and input. Error bars indicate SD from three biological replicates. ***P < 0.005, two-tailed Student’s t test. b The indicated cell populations were subjected to luciferase assays using Enhancer E. Error bars indicate SD from three biological replicates. ***P < 0.005, two-tailed Student’s t test. c Volcano plots of RNA-Seq analyses in MCF10A cells transduced with the indicated vectors. The data was compared and normalized to control cells (Empty-OE or non-targeting sgRNA). Dark dots are the genes that were significantly upregulated or downregulated in RNA level compered to control with cutoff of absolute LFC > 0.5 and adjusted p value < 0.05. YAP is labeled green and TRAM2 red. d Venn diagrams representing the intersection of proteins that were upregulated in MCF10A-TRAM2 and MCF10A-YAP^5SA cells (raw p value < 0.05 and log2 fold-change > 0.7) and were downregulated in MCF10A-YAP^5SA cells upon TRAM2-KO1 and TRAM2-KO2 (raw p value < 0.05 and log2 fold-change < − 0.7). e Representative overview images of Matrigel-coupled wound-healing invasion assays of MCF10A-TRAM2 cells transduced with the indicated vectors at 0 and 48 h. f Wound confluence was calculated using IncuCyte system based on 4 independent Matrigel-coupled wound-healing invasion assay experiments, as in e. g In vitro migration assays of MCF10A-TRAM2 cells transduced with the indicated lentiviral vectors. Cells were labeled with GFP for tracking cells under the microscope. Error bars indicate SD from counted cells in each condition (NT: n = 7303; FSTL1 KO1: n = 7051; FSTL1 KO2: n = 5368). ***P < 0.005, two-tailed Student’s t test