Fig. 1: EGFR/MAPK/ERK signaling and associated gene regulation.

a Schematic of the various techniques applied in this study to evaluate copper stimulated transcriptional/genomic dynamics, with the canonical EGFR/MAPK/ERK signaling pathway. Figure 1/panel a created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license. b A549 KAS-seq gene body volcano plots (Cu-supplemented vs. unsupplemented) 15 min post-cell treatment. Significantly upregulated genes in red; horizontal gray lines denote p = 0.05; n = 2 biological replicates for each condition. Low Cu and High Cu conditions correspond to 10 and 20 µM CuCl₂ supplementation, respectively. p-value denotes two-sided Wald test as calculated by DESeq2, without correction for multiple observations. c Changes in HEK 293 T phosphorylation and/or total protein levels from proteome profiler phospho-kinase antibody arrays following 10 min, 20 µM CuCl₂ treatment (vs. untreated). n = 3 biological replicates for each condition. p-value denotes two-sided t-test without correction for multiple observations. d RNA-qPCR fold-changes of select transcripts following copper supplementation for indicated times into A549 cells. Box plot values defined as follows: box boundaries (median of first quartile to median of third quartile) and central horizontal line (median), mean expression value shown on the plot as an “x”, whisker boundaries define 1.5 times the interquartile range. Data points correspond to biological replicates, p-values calculated as paired two-tailed t-test. Sample sizes were as follows for each transcript at 60 min, 120 min, and 240 min post-Cu; EGR1 (n = 2, 6, 10), NAB1 (n = 2, 6, 6), NAB2 (n = 2, 6, 6), CTR1 (n = 2, 2, 10–22). Source data are provided as a Source Data file, as are exact p-values. * denotes p < 0.05, ** denotes p < 0.01, and *** denotes p < 0.001.