Figure 1.
Figure 1. SOX11 directly regulates CXCR4 and PTK2 gene expression. (A) GSEA analysis on gene expression profiling microarray data from SOX11+ (Z138CT) and SOX11KD (Z138KD) xenograft tumors and tissue SOX11+ and SOX11− MCL primary cases, using microenvironment-related gene sets described in “Methods.” Normalized enrichment score (NES), P value, and false-discovery rate (FDR) are shown. FDR <0.2 indicates statistical significance. (B) ChIP-qPCR enrichment in Z138 and JVM2 MCL cell lines of the SOX11 pulldown in CXCR4 and PTK2 loci (peak 1 and peak 2; supplemental Figure 1). DNA enrichment is displayed as fold change relative to their respective input chromatin. (C) Luciferase assays in transient cotransfections of CXCR4 and PTK2 enhancer GL4.23Luc (peak 1 and peak 2) with full-length SOX11 (pcDNA3SOX11) and truncated SOX11 (pcDNA3 ∆HMGSOX11) in HEK293T cell line. Results are shown as percentage fold induction referred to luciferase activity in cotransfection with the empty vector (pcDNA3-Ø). Red arrows indicate the most significant activated enhancers by SOX11 expression. (D) Western blot experiments showing total cellular FAK and SOX11 protein levels in Z138CT, Z138KD, JVM2CT, and JVM2SOX11+ MCL cell line models (“Methods”). Notice that the slight band shift of SOX11 in JVM2SOX11+ is a result of its expression with a FLAG tag. Actin was used as a loading control (left). Bar graph representing fold change differences in percentage of FAK protein levels in MCL cell lines, corrected by quantification of actin expression levels. Relative fold enrichment is displayed in reference to FAK protein levels of the SOX11+ Z138CT cell line (right). (E) Bar graph representing the mean fluorescence of FC experiments showing the expression levels of cellular surface CXCR4 protein levels in Granta519 and Z138CT vs its KD counterparts and JVM2SOX11+ vs JVM2CT MCL cell lines. Granta519 was used as a CXCR4− control cell line. Isotype immunoglobulin G control antibody was used as a negative CXCR4 control staining. Average levels of the PTK2 (F) and CXCR4 (G) probe set signals obtained from the analysis of Affymetrix HG-U133 2.0 plus microarrays from our 34 PB primary MCL tumors (14 SOX11+ and 20 SOX11−). Bar plot represents the mean percentage ± standard deviation of 3 independent experiments. The significance of difference was determined by independent samples Student t test: *P < .05, **P < .01, ***P < .001. WT, wild type.

SOX11 directly regulates CXCR4 and PTK2 gene expression. (A) GSEA analysis on gene expression profiling microarray data from SOX11+ (Z138CT) and SOX11KD (Z138KD) xenograft tumors and tissue SOX11+ and SOX11− MCL primary cases, using microenvironment-related gene sets described in “Methods.” Normalized enrichment score (NES), P value, and false-discovery rate (FDR) are shown. FDR <0.2 indicates statistical significance. (B) ChIP-qPCR enrichment in Z138 and JVM2 MCL cell lines of the SOX11 pulldown in CXCR4 and PTK2 loci (peak 1 and peak 2; supplemental Figure 1). DNA enrichment is displayed as fold change relative to their respective input chromatin. (C) Luciferase assays in transient cotransfections of CXCR4 and PTK2 enhancer GL4.23Luc (peak 1 and peak 2) with full-length SOX11 (pcDNA3SOX11) and truncated SOX11 (pcDNA3 ∆HMGSOX11) in HEK293T cell line. Results are shown as percentage fold induction referred to luciferase activity in cotransfection with the empty vector (pcDNA3-Ø). Red arrows indicate the most significant activated enhancers by SOX11 expression. (D) Western blot experiments showing total cellular FAK and SOX11 protein levels in Z138CT, Z138KD, JVM2CT, and JVM2SOX11+ MCL cell line models (“Methods”). Notice that the slight band shift of SOX11 in JVM2SOX11+ is a result of its expression with a FLAG tag. Actin was used as a loading control (left). Bar graph representing fold change differences in percentage of FAK protein levels in MCL cell lines, corrected by quantification of actin expression levels. Relative fold enrichment is displayed in reference to FAK protein levels of the SOX11+ Z138CT cell line (right). (E) Bar graph representing the mean fluorescence of FC experiments showing the expression levels of cellular surface CXCR4 protein levels in Granta519 and Z138CT vs its KD counterparts and JVM2SOX11+ vs JVM2CT MCL cell lines. Granta519 was used as a CXCR4− control cell line. Isotype immunoglobulin G control antibody was used as a negative CXCR4 control staining. Average levels of the PTK2 (F) and CXCR4 (G) probe set signals obtained from the analysis of Affymetrix HG-U133 2.0 plus microarrays from our 34 PB primary MCL tumors (14 SOX11+ and 20 SOX11−). Bar plot represents the mean percentage ± standard deviation of 3 independent experiments. The significance of difference was determined by independent samples Student t test: *P < .05, **P < .01, ***P < .001. WT, wild type.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal