Fig. 1: Generation of BAP1-KO cell clones and effect of BAP1 loss on the gene expression in human mesothelial cells.
From: CAMK2D: a novel molecular target for BAP1-deficient malignant mesothelioma
a Generation of BAP1‐KO cell lines using the CRISPR/Cas9 system using a single guide RNA sequence against exon 4 of the BAP1 gene locus. b Establishment of BAP1-KO cell clones using human mesothelial cell lines, MeT‐5A and HOMC-D4. BAP1 protein expression was determined by Western blot analysis using GAPDH as the internal control. c Gene expression analysis represented by heatmap of upregulated (seven genes; fold change, >3) and downregulated (40 genes, fold change, <0.1) genes in BAP1‐KO cells (#1 and #2) compared with parental (P) and BAP1-WT (Ctrl) cells. The heatmap was constructed based on the normalized values of all samples using TreeView (Cluster 3.0) http://jtreeview (http://jtreeview.sourceforge.net). The corresponding upregulated or downregulated genes in the heatmap are shown on the right side. d RT-PCR analysis for mRNA expression levels of upregulated or downregulated genes in the MeT-5A and HOMC-D4 cells. Representative agarose gels for the RT-PCR products from the parental cells (P), BAP1-WT cells (Ctrl), and BAP1-KO cell lines (#1 and #2) are shown.
