Supplemental Figures and Tables from Caspase-9b Interacts Directly with cIAP1 to Drive Agonist-Independent Activation of NF-κB and Lung Tumorigenesis

Abstract

<p>Supplemental Figure 1: Larger legend table for symbols in fig 1A. Supplemental Figure 2: The expression of BIRC5, CDKN1A, CXCL1 and IER3 correlates with expression of C9b, but not C9a, in human NSCLC tissues. Supplemental Figure 3: Down-regulation of RIP1 abolishes the effect of C9b on IκBα level.. Supplemental Figure 4: The role of C9b in the NF-κB pathway is translational to other cell lines, H838 and HBEpC. Supplemental Figure 5: C9b activates the canonical NF-κB pathway in H838 cells and HBEpCs. Supplemental Figure 6: C9b level inversely correlates with IκBα level in NSCLC cells. Supplemental Figure 7: Down-regulation of cIAP2 has no effect on NF-κB activation and does not inhibit the effect of C9b on the NF-κB pathway. Supplemental Figure 8: Cleavage of C9b is not important for interaction with cIAP1. Supplemental Figure 9: Tumors formed from NSCLC cells stably expressing WT C9b have higher Ki-67 expression (cell proliferation) than tumors formed from control cells or cells expressing IBM-mutated (Mut) C9b. Supplemental Figure 10: The interaction with cIAP1 mediates the enhancing effect of C9b on the cell viability and survival/AIG of NSCLC cells and HBEpCs. Supplemental Figure 11: Suppression of the canonical NF-κB pathway by ectopically expressing the suppressor, IκBα, increases NIK level. Supplemental Figure 12: Down-regulation of NIK does not affect the survival/AIG of NSCLC cells. Supplemental Table 1: qPCR primers for NF-κB target genes utilized in the study Supplemental Table 2: List of NF-κB-related genes that expression was affected by down-regulation of C9b in A549 cells or up-regulation of C9b in HBEpC cells (identified by deep RNA sequencing). Supplemental Table 3: List of NF-κB target genes that expression is contrastingly affected by down-regulation of C9b in A549 cells versus by up-regulation of C9b in HBEpCs. Supplemental Table 4: NSCLC cell lines used in Figure 2D or Supplemental Figure 6. Supplemental Table 5: Sequences of negative control si-RNA and caspase-9a or -9b si-RNA utilized in the study.</p>