Fig. 1: Nuclear EGFR drives a unique transcriptional profile.

a nEGFR tracking in T47D EGFR-YFP cells. T47D cells expressing the EGFR-YFP fusion protein were serum starved overnight, then treated with 100 ng/ml of EGF, 10 μM of cSNX1.3, or 10 μM of the control peptide cPTD4 and imaged every 10 min for 24 h. Images were quantified via MATLAB. Data represents the mean ± SEM of the integrated intensity of EGFR-YFP in the nucleus of T47D cells, ±SEM. Dash line represents the time of drug added at 30 min. cSNX1.3 treatment is statistically different from cPTD4 and EGF treatments; p < 0.0001. Statistics were analyzed using one-way ANOVA, Dunnett’s post hoc. b Cell viability after cSNX1.3 and erlotinib treatment. MDA-MB-468 cells were treated with the drug treatments shown above for 72 h. Cell viability was determined with an MTT assay and normalized to vehicle control. Data represents the percentage of cell viability ± SEM of 3 biological replicates. Statistics were analyzed via one-way ANOVA, Dunnett’s post hoc; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, compared to cSNX1.3 alone. The Bliss independence model was used to test for synergy, resulting in a score of 3.499. c Heatmap; MDA-MB-468 cells were serum starved for 12 h, stimulated with 10 ng/mL EGF, and treated with cSNX1.3 (10 μM) or erlotinib (15 μM) for 12 h. Samples were collected and next generation sequencing was performed by GENEWIZ from Azenta Life Sciences. Significantly differentially expressed (P_adj < 0.05 and abs(log₂FC)>1) genes in the cells treated with cSNX1.3 + EGF (referred to as cSNX1.3) are visualized using their z-score across three different conditions (cSNX1.3, erlotinib, and EGF control). Venn diagram; number of significant (P_adj < 0.05) differentially expressed genes shared by both cSNX1.3 and erlotinib treatments. d Functional enrichment in the significant (P_adj < 0.05) differentially expressed genes between cSNX1.3 and erlotinib treatments. GO term enrichment analysis of cSNX1.3 with log₂FC > 1 (top, left). GO term enrichment analysis of erlotinib with log₂FC > 1 (top, right) and log₂FC < -1 (bottom). e Natural killer cell-mediated cytotoxicity genes regulated by cSNX1.3 or erlotinib. Log₂FC of significant (P_adj < 0.05) genes associated with natural killer cell-mediated cytotoxicity in cSXN1.3 and erlotinib. The human gene set used for identifying natural killer cell-mediated cytotoxicity genes can be found under the standard name KEGG_NATURAL_KILLER_CELL_MEDIATED_CYTOTOXICITY.