![TRAF6-dependent and -independent gene expression profiling (GEP) of oncogenic CARD11. (A) Comparative gene set enrichment analysis of 3 ABC DLBCL–derived NF-κB gene signatures of CARD11 L232LI in parental vs TRAF6 KO BJAB cells. Three different NF-κB signatures were compared, with q values for the individual signatures shown in the figure. (B) Heat map of genes differentially expressed in parental and TRAF6 KO BJAB cells expressing CARD11 WT or L232LI (log fold change [LFC] > 1.3; adjusted P value [Padj] < .01). Genes are ordered based on the Padj values of parental CARD11 L232LI vs CARD11 WT. Red denotes higher, blue denotes lower expression on the color scale; n = 5 for each sample. Genes highlighted in blue are induced in the absence of TRAF6. (C) Signed Padj plot comparing gene induction by CARD11 L232LI in parental vs TRAF6 KO BJAB cells. Transcripts enriched in TRAF6 KO cells are depicted in blue. (D) Evaluation of the expression of selected transcripts by qRT-PCR in parental and TRAF6 KO BJAB cells transduced with CARD11 WT or CARD11 L232LI. Changes in transcript expression after MALT1 protease inhibition by MLT-748 (2 μM; 18 hours) in CARD11 L232LI–expressing TRAF6 KO BJAB cells. n = 5, all error bars depict the mean ± SD; left panel: ANOVA with Tukey multiple comparisons; right panel: unpaired Student t test; ∗∗P < .01; ∗∗∗P < .001; and ∗∗∗∗P < .0001. (E) Western blot analysis of BJAB and TRAF6 KO BJAB cells expressing CARD11 WT and L232LI to determine cleavage of Regnase-1 and Roquin-1/2 and expression of IκBNS. Unspecific bands are marked with an asterisk.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/142/23/10.1182_blood.2023021299/1/blood_bld-2023-021299-gr2de.jpeg?Expires=1719277216&Signature=yFdlwC1Rsi00B9dRtQBc9eJAuv5wwywexvO2WbzC~iLw1PvIW8emWUi4Lq7wqlg~TWIt0x4o3SWfCLEQjiYxtYwUIxuKufPs5gsvtsA-OszRw7QoReRW00TIJVmIvU4mOCjs9IROYA7xnSn6JmeZ2szTruc5W1EI4HOuqN9WxFJAsX9dp~~2Xd4EVWPh3zjHRgg8rwzUZJEkp6Y8GVrOfF2WHz7lqL-3diFpTr479SyLC5yzUACqQHUmM3WhE950pSnfv8R5PP6GKGrfe5h-Ax5kJZYKDhLubQYReWc49NdHct0pVtX0fp6ffBV3zimKOK~JccKYkbOZizq-mfEsqw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
TRAF6-dependent and -independent gene expression profiling (GEP) of oncogenic CARD11. (A) Comparative gene set enrichment analysis of 3 ABC DLBCL–derived NF-κB gene signatures of CARD11 L232LI in parental vs TRAF6 KO BJAB cells. Three different NF-κB signatures were compared, with q values for the individual signatures shown in the figure. (B) Heat map of genes differentially expressed in parental and TRAF6 KO BJAB cells expressing CARD11 WT or L232LI (log fold change [LFC] > 1.3; adjusted P value [Padj] < .01). Genes are ordered based on the Padj values of parental CARD11 L232LI vs CARD11 WT. Red denotes higher, blue denotes lower expression on the color scale; n = 5 for each sample. Genes highlighted in blue are induced in the absence of TRAF6. (C) Signed Padj plot comparing gene induction by CARD11 L232LI in parental vs TRAF6 KO BJAB cells. Transcripts enriched in TRAF6 KO cells are depicted in blue. (D) Evaluation of the expression of selected transcripts by qRT-PCR in parental and TRAF6 KO BJAB cells transduced with CARD11 WT or CARD11 L232LI. Changes in transcript expression after MALT1 protease inhibition by MLT-748 (2 μM; 18 hours) in CARD11 L232LI–expressing TRAF6 KO BJAB cells. n = 5, all error bars depict the mean ± SD; left panel: ANOVA with Tukey multiple comparisons; right panel: unpaired Student t test; ∗∗P < .01; ∗∗∗P < .001; and ∗∗∗∗P < .0001. (E) Western blot analysis of BJAB and TRAF6 KO BJAB cells expressing CARD11 WT and L232LI to determine cleavage of Regnase-1 and Roquin-1/2 and expression of IκBNS. Unspecific bands are marked with an asterisk.
