Figure 6.

In vitro CRISPR/Cas9 knockout resulted in a more proliferative and chemoresistant cellular phenotype. (a) Endogenous expression of Arid1a in osteosarcoma cell lines as demonstrated by immunofluorescence staining showing nuclear localization of Arid1a (Actin = Red, Nuclei = Blue). (b) Schematic for targeting Arid1a with CRISPR/Cas9. (c) Relative mRNA expression of Arid1a after knockout in two biological replicates of U2-OS and SJSA-1. The data represent an average of three technical replicates and error bars represent the standard error of the mean (** p-value < 0.05; *** p-value < 0.005; **** p-value < 0.0005). (d) Real-time cell proliferation assay for 48 h in U2-OS cell lines, error bar represents the standard error of the mean (**** p-value < 0.0001; individual t-tests compared between groups at different time points), (n = 5). (e) Real-time cell migration assay at different time points (T1 = 0 h, T5 = 24 h), error bars represents the standard deviation of mean (**** p-value < 0.0001), (n = 3). (f) Doxorubicin chemosensitivity assay at 72 h, IC₅₀ for KO U2-OS = 0.7 μM, and Control U2-OS 0.3 μM.