*3.6. Validation of RNA-Seq Gene Expression Levels Using RT-qPCR*

To validate the accuracy of RNA-Seq results, ten genes in HL60 and KG1a were also detected by RT-qPCR. As a result, the RNA-Seq-detected expression regulation of 20 genes were verified by RT-qPCR detection (Figure 7A,B). Additionally, the qPCR-detected CD38 expression (Figure 1E) was also in agreement with that detected by RNA-Seq (Figure 3B). These data indicated that the RNA-Seq results were reliable.

**Figure 7.** Validation of RNA-Seq DEGs using RT-qPCR: (**A**,**B**) the expression levels of 10 selected DEGs in the iron nanoparticle-treated HL60 (**A**) and KG1a (**B**) cells. All values are mean ± SD with *n* = 3. ns, no significance; \*, *p* < 0.05; \*\*, *p* < 0.01, \*\*\*, *p* < 0.001, \*\*\*\*, and *p* < 0.0001. (**C**,**D**) The comparison of fold change detected by RT-qPCR and RNA-Seq in the iron nanoparticle-treated HL60 and (**C**,**D**) KG1a cells.
