Figure 1.

High dose zinc aspartate negatively regulates genes controlling cell cycle entry and progression in CD4⁺ T cells. (A) Principal component analysis (PCA) of the individual biological samples used for bulk RNA sequencing. (B) Unsupervised clustering heatmap showing differentially expressed genes (DEG) between individual biological samples. (C) TOP Gene ontology (GO) Biology Function pathways based on p-value. Shown is the –log₁₀ of the p-values of the individual pathways. (D) TOP KEGG pathways based on p-value. Shown is the –log₁₀ of the p-values of the individual pathways. (E) Normalized gene expression of Mcm2, Mcm5, Mcm6, Cdk2, E2f1, E2f3, and E2f6 determined by RNA sequencing. Statistical analysis in (E) by unpaired student’s t test. Dots represent individual mice used to isolate OT-II TCR tg CD4⁺ T cells. RNA sequencing data was generated in biological triplicates from 3 mice. KEGG was used for pathway analysis²⁶. p* < 0.05, **p < 0.01, ***p < 0.001.