Figure 1

Generation of MAPK4-KO HCT-8 cells. (A) Schematic image of the strategy to generate MAPK4-KO HCT-8 cells and genotyping. Two gRNAs were designed to hybridize and eliminate the MAPK4-coding site in HCT-8 cells. Genomic DNA of each cell was extracted and analyzed by PCR analysis. PCR of P1-4 site was not amplified with WT HCT-8 cells because of long introns in-between. The sequence of the P1-4 product in MAPK4-KO HCT-8 cells is shown in the image. (B) Cell growth assay of WT and MAPK4-KO HCT-8 cells. No significant difference was observed with cell types (Two-way ANOVA, p = 0.1316). (C) LDH-based cell mortality of WT and MAPK4-KO HCT-8 cells under non-infected conditions. Percentages represent the ratio of LDH release to whole cell lysates. No significant difference was observed (Student’s t test, p = 0.1393). All graphs show the mean ± SEM for more than three independent experiments. All images are representative of three independent experiments.