Fig. 2: CASP4 knockout decreases angiogenesis and expression of cell migration-related genes.

a Representative images of immunoblotting analysis of CASP4 protein in mock and CASP4-knockout (C4KO) NCI-H292 cells. b Representative images of fluorescent immunostaining of CASP4 protein in mock and CASP4-knockout (C4KO) cells. Scale bar, 20 μm. c Sequence showing insertion and deletion in exon 2 of human CASP4 in C4KO NCI-H292 cells. d Flowchart of the RNA-seq experiment and data analysis. e The top eight downregulated GO terms in NCI-H292 cells following CASP4 KO based on GO analysis of CDS expression data obtained from RNA sequencing. The horizontal axis shows accumulative hypergeometric p values. f Representative heatmap of cell migration- and angiogenesis-related genes downregulated by CASP4 KO. g Differentially expressed genes that overlapped between C4OE upregulated and C4KO downregulated in “angiogenesis” and “positive cell migration. h Positive regulation of cell migration- and angiogenesis-related mRNAs, such as EPHA2 and PTGS2, based on RT-qPCR in C4OE and mock cells. i Positive regulation of cell migration- and angiogenesis-related mRNAs, such as EPHA2, and PTGS2, based on RT-qPCR in C4KO and mock cells.” N.S: nonspecific. Data are expressed as mean ± standard deviation (n = 3). **P < 0.01 compared with mock. Densitometry values are shown above each blot.