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. 2022 Mar 1;23(5):2732. doi: 10.3390/ijms23052732

Figure 4.

Figure 4

ARMC4 expression decreased NF-κB target gene expression. (A) An RNA-seq was conducted in the presence or absence of 10 ng/mL of IL-1β in HEK293 cells. Overexpression of ARMC4 decreased 56.3% (224 genes) of NF-κB target expression by 2-fold or more (ARMC4 + IL-1β/Ctrl + IL-1β ≤ 0.5), while leaving 39.2% (156 genes) unaffected (ARMC4 + IL-1β/Ctrl + IL-1β = 0.5–2), and merely 4.5% (18 genes) were increased by 2-fold or more (ARMC4 + IL-1β/Ctrl + IL-1β ≥ 2). Cells were treated or untreated with IL-1 β for 4 h before RNA was collected. (B–D) Ingenuity pathway analysis (IPA). IPA analysis identified near 80% of ARMC4-downregulated NF-κB target genes are cancer related (B), suggesting ARMC4’s important negative NF-κB regulation function in cancer. IPA also identified that ARMC4 has several top network cellular functions, including cell-cell signaling, proliferation, and cell morphology (C). Furthermore, IPA analysis identified a typical network that was regulated by ARMC4, in which NF-κB is at the hub of this network (D). (E) qPCR analysis of typical genes identified, confirming NFKBIA, IL-8, and CCL20 are NF-κB-inducible genes (+IL-1β condition) and were significantly downregulated by the overexpression of ARMC4 in both HEK293 (top row) and HT29 cells (bottom row) under IL-1β treatment. The data represent the means ± SD from three independent experiments. * p < 0.05 Ctrl + IL-1β vs. Ctrl group; $ p < 0.05 ARMC4 + IL-1β vs. Ctr + IL-1β group; n = 3.