## 3 Results
### Mapping and Identification of DEGs  
Raw data were mapped onto the human reference genome (GRCh38) using the BBMap program with default parameters. The proportion of mapped reads in each library ranged from 90.7% to 89.9% (Figure 2). Approximately 10% of the sequenced reads in each library were not associated with human transcripts.  
\begin{table}
\begin{tabular}{c c c c c} \hline
**Index** & **Condition** & **Treatment** & **Library Name** & **Replicate** \\ \hline
1 & Control & Distilled water & DW-1 & 1 \\
2 & Control & Distilled water & DW-2 & 2 \\
3 & Control & Distilled water & DW-3 & 3 \\
4 & GP-P & GP extract from plants & GP-P1 & 1 \\
5 & GP-P & GP extract from plants & GP-P2 & 2 \\
6 & GP-P & GP extract from plants & GP-P3 & 3 \\
7 & GP-C & GP extract from callus & GP-C1 & 1 \\
8 & GP-C & GP extract from callus & GP-C2 & 2 \\
9 & GP-C & GP extract from callus & GP-C3 & 3 \\
10 & GP-CR & GP extract from callus treated with radiofrequency & GP-CR1 & 1 \\
11 & GP-CR & GP extract from callus treated with radiofrequency & GP-CR2 & 2 \\
12 & GP-CR & GP extract from callus treated with radiofrequency & GP-CR3 & 3 \\ \hline \end{tabular}
\begin{tabular}{c c c} \hline
**Index** & **Condition** & **Treatment** & **Library Name** & **Replicate** \\ \hline
1 & Control & Distilled water & DW-1 & 1 \\
2 & Control & Distilled water & DW-2 & 2 \\
3 & Control & Distilled water & DW-3 & 3 \\
4 & GP-P & GP extract from plants & GP-P1 & 1 \\
5 & GP-P & GP extract from plants & GP-P2 & 2 \\
6 & GP-P & GP extract from plants & GP-P3 & 3 \\
7 & GP-C & GP extract from callus & GP-C1 & 1 \\
8 & GP-C & GP extract from callus & GP-C2 & 2 \\
9 & GP-C & GP extract from callus & GP-C3 & 3 \\
10 & GP-CR & GP extract from callus treated with radiofrequency & GP-CR1 & 1 \\
11 & GP-CR & GP extract from callus treated with radiofrequency & GP-CR2 & 2 \\
12 & GP-CR & GP extract from callus treated with radiofrequency & GP-CR3 & 3 \\ \hline \end{tabular}
\end{table}
Table 1: List of the 12 HaCaT samples used for RNA sequencing.  
Figure 1: Generation and cultivation of GP cells from GP leaves using tissue culture and bioreactors. (**A**) GP plant used for tissue culture. (**B**) Sterilization of the GP leaves. (**C**) Cutting of the GP leaves. (**D**) Transfer of the GP leaves on the plate containing MS medium. (**E**) Induction of the GP callus. (**F**) Propagation of the GP callus. (**G**) Suspension cell cultivation of the GP callus in liquid medium. (**H**) Mass cultivation of the GP cells in a bioreactor. GP, _Gymostemma pentaphyllum_; MS, Murashige and Skoog.  
We identified DEGs by comparing the GP extract-treated and control samples. The numbers of DEGs identified under treatments with the three extracts were very similar (Figure 3). Among the extracts, GP-C (93 DEGs) showed the highest number of DEGs, followed by GP-CR (90 DEGs) and GP-P (87 DEGs). In addition, we combined all GP extract-treated samples, referred to as GP-all, and compared them to the control conditions, resulting in the identification of 176 DEGs. In both GP-P and GP-all conditions, the number of upregulated genes was higher than that of downregulated genes. The number of upregulated genes (125 genes) was almost three times that of the downregulated (51 genes) in GP-all. In contrast, the number of downregulated genes was much higher than that of the upregulated genes in the GP-C and GP-CR conditions.