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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Cancer Immunol Res. 2019 Oct 17;7(11):1760–1774. doi: 10.1158/2326-6066.CIR-18-0832

Figure 6. Effects of knocking down IKKs, TNFRSF1A, and NF-κB family members altered gene expression and cell proliferation in UM-SCC 1 cells.

Figure 6.

(A) CHUK (IKKα), IKKβ, IKKγ were knocked down with two siRNA each in UM-SCC1 cells for 48 hours, then treated with TNF-α (10ng/ml) or LTβ (100ng/ml) 6 hours prior to harvesting. Relative gene expression of RELB, NF-κB2 and BIRC3 were verified by qRT-PCR in triplicates compared to cells transfected with control siRNA without treatment. The data were calculated as mean ± SD. * indicates statistical significance compared to no treatment, and # indicates statistical significance compared to control siRNA with same treatment (student t-test, P-value≤0.05). The two siRNAs were transfected independently and served as the biological replicates. (B) Cell density was measured after gene knockdown in UM-SCC 1 cell line in 6 replicates. XTT assay was performed 72 hours after siRNA transfection, without treatment (grey), with stimulation by TNF-α for final 16 hour (dash), or by LTβ for final 24 hour (black) before adding XTT reagents. White bar indicates the control condition without siRNA transfection. The data are from one representative siRNA knockdown of two repeated experiments, and statistical significance was calculated from six replicates (t test, P<0.05).