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. 2022 Apr 8;12:5944. doi: 10.1038/s41598-022-09963-7

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© The Author(s) 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Functional relevance of NFκB binding motifs in human hnRNPD promoter. (A) Location and nucleotide sequence of the four putative NFκB (RelA) binding motifs as identified by in silico analysis of hnRNPD promoter. Each binding motif was individually subjected to site directed mutagenesis in promoter reporter construct pVKS-1 and the resulting constructs were named as pVKS-1 Mut1, pVKS-1 Mut2, pVKS-1 Mut3 and pVKS-1 Mut4 respectively. Mutation in each motif was confirmed by double stranded DNA sequencing and the mutated motifs have been shown in bold face along with their electrophoretogram. (B) Luciferase activity in SCC-4 cells after transfection with each mutant construct was assayed and compared with the activity of pVKS-1. Other details are given in Materials and Methods. Values are mean ± SD from four independent experiments performed in triplicate. Results were analyzed using a paired two tailed Student’s t-test and values significantly different from pVKS-1 marked by (*P ≤ 0.05).