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. 2022 Oct 21;13(10):885. doi: 10.1038/s41419-022-05308-4

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Immunoblotting analysis showing TNFR1 levels in KAL^LU+ cells treated with TNFR1 siRNA (si-TNFR1) or si-Control. β-actin, a protein-loading control. b Growth curve of KAL^LU+ cells treated with si-TNFR1 or si-Control. Mean ± SD (three repeats); ***P < 0.001; Two-way AVOVA statistical test. c Immunoblotting analysis shows indicated protein levels in KAL^LU+ cells treated with si-Control (si-Con) or si-TNFR1. β-actin, a protein-loading control. d, e Comparing tumor sizes (d) and numbers (e) derived from KAL^LU+ cells treated with si-Control (si-Con) or si-TNFR1 in WT and KA/KA mice (recipient, n = 5/group). Each symbol represents a tumor. Mean ± SD; ***P < 0.001; Student’s t-test. f H&E-stained and E-cadherin IHC-stained sections of tumors derived from KAL^LU+ cells treated with si-Control or si-TNFR1 in KA/KA mice (recipient). Scale bar: 40 μM. g K5 IHC staining for lung SCCs derived from si-TNFR1 KAL^LU+ cells in KA/KA mice at 1.5 months after cell injections. Scale bar: 50 μM. h Immunoblotting analysis showing the levels of K5 and TNFR1 in KAL^LU+ cells treated with si-Control and si-TNFR1. β-actin, a protein-loading control. i RT-PCR analysis showing TNF expression in WT and L-KA/KA lungs (left) and in M2C and KAL^LU+ cells (right) treated with si-Control (si-Con) and si-TNFR1 (n = 3/group). Mean ± SD (three repeats per group); ***P < 0.001; Student’s t-test. j, k Comparing tumor sizes (j) and tumor numbers (k) derived from KAL^LU+ cells treated with si-Control (si-Con) or si-TNF in WT and KA/KA mice (n = 5/group). Mean ± SD; *P < 0.05; **P < 0.01; ***P < 0.001; Student’s t-test. l Weights of tumors isolated from nude mice receiving subcutaneous injections of KAL^LU+ cells treated with si-Control (si-Cont) and si-TNF RNA (n = 5/group). Mean ± SD; **P < 0.01; Student’s t-test.