Fig. 6

Lamin B1 is essential for EMT. a Principle component analysis (PCA) plot of RNA-seq based expression profiling in KD conditions. Each time point was normalized to its respective control. b Number of genes that were differentially expressed (DE) upon lamin B1 knockdown (KD) during the EMT process. Inside each bar, the proportion of genes that are direct targets of lamin B1 in the ChIP-seq experiment of NMuMG cells untreated (blue) or treated with TGF-β for 8 h (orange) or 24 h (red) are shown. c Venn diagram representing the overlap between upregulated genes after 8 h of TGF-β treatment in KD and C conditions. d Western blotting showing protein levels of different EMT markers and lamin B1 in C and KD NMuMG cells that were untreated or treated with TGF-β for 8 or 24 h. Tubulin was used as a loading control. Images are representative of three independent replicates. e Migration (left panel) and invasion (right panel) assays performed with C and KD NMuMG cells after 24 h of TGF-β treatment. Data are presented as mean ± SD, n = 3. f Chip-qPCR of three selected EMT-related lamin B1 + regions at 8 h after TGF-β treatment in C and KD conditions. One of the selected regions from Fig. 4d was used as a negative control (prOCT4). Data from qPCR were normalized to the input and expressed as the fold-change relative to the untreated C condition, which was set as 1. Data are shown as mean ± SD; n = 3. g Western blotting of overexpressed GFP-control or GFP-hLB (human LB1) from C or KD NMuMG cells. Tubulin was used as a loading control (left panel). Images are representative of three independent replicates. Migration assay of C and KD NMuMG cells transfected with either GFP-control or GFP-hLB (right panel). Data are shown as mean ± SD; n = 2