Fig. 3

CASC15 expression is strongly correlated with and regulates SOX4 expression. a-b Correlation between CASC15 and SOX4 expression in various B-ALL and AML cell lines. c-d RT-qPCR of CASC15 and SOX4 in REH cells (c) and RS4;11 (d) following knockdown of CASC15 using three independent siRNA sequences. Statistically significant differences from control are noted as follows: p < 0.05 (*); p < 0.01 (**); p < 0.0005 (***). e-f Western blot analysis of SOX4 protein levels in RS4;11 (e) and REH (f) cell lines upon CASC15 knockdown. (g) RT-qPCR of CASC15 and SOX4 in REH cells following CRISPR/Cas9 mediated targeting. Statistically significant differences from control are denoted as follows: p < 0.05 (*); p < 0.01 (**). h RT-qPCR of murine Casc15 and Sox4 on RNA extracted from FACS-purified hematopoietic progenitor fractions shows a rough correlation in their expression, particularly in the B-cell subsets. i RT-qPCR for murine Sox4 following transduction of bone marrow cells with murine Casc15 shows upregulation of expression. All qPCR analyses for CASC15 were performed with primer set #1 and normalized to GAPDH or actin, except where otherwise noted. Experiments were repeated three times for validation. Experiments were repeated three times for validation