*3.5. Gene Expression Overview*

As shown in Figure 4A, a total of 15,721 genes were identified through RNA sequencing, and 14,193 genes were co-expressed in the Ctr, L4, and M21 samples. A principal component analysis (PCA) was performed to assess the transcriptomics of the different samples. Figure 4B reveals that the Ctr, L4, and M21 samples were well divided into three characteristic groups by PCA, which indicated that there were significant differences in the

transcriptomics between the groups [41]. Further analysis indicated that compared to the Ctr samples, 121 genes (81 upregulated, 40 downregulated) and 30 genes (17 upregulated, 13 downregulated) were significantly differentially regulated in the L4 and M21 samples, respectively (Figure 4C,D, Tables S2 and S3). The results indicated that DMM can significantly interfere with the gene expression of Jurkat T cells, even at a low dose (4.12 mg/L L4 group). This result is consistent with that of the toxicity test shown in Figure 1. Previous studies showed that the highest dimethomorph residue was 6.8 mg/kg for leafy vegetables and stalk and stem vegetables and 6.11 mg/kg for *Dendrobium officinale* [42,43]. Considering the cytotoxicity and transcriptomic results of our study, the immunotoxicological effects of DMM should be emphasized.

**Figure 3.** RNA electropherogram (**A**) and sample correlations based on RNA sequencing (**B**). (**A**): M, marker; 1–3, EC50 group; 4–6, EC25 group; 7–9, EC10 group; 10–12, control group.
