## 3 Results
### DEGs in Response to GP Extracts  
We selected the top ten significant DEGs in each condition based on the Manhattan distance (Figure 4). Among the 87 DEGs in GP-P (Table S1), the top ten DEGs comprised five significantly downregulated genes (_CARD17_, _LCN12_, _FUOM_, _LOC105374145_, and _LOC107984639_) and five significantly upregulated genes (_KCND3_, _LOC101928092_, _NLRP6_, _S100A8_, and _ELDR_) (Figure 4A). In addition, we identified five long intergenic non-coding (LINC) RNAs _LINC00539_, _LINC01103_, _LINC01765_, _LINC01792_, and _LINC02355_. We identified 93 DEGs after treatment with GP-C (Table S2). The number of downregulated genes (62 genes) was twice that of the upregulated genes (31 genes) (Table S2). Similarly, among the top ten significant DEGs identified, the number of significantly downregulated genes (seven genes) was higher than that of the upregulated genes (three genes) (Figure 4B). The top downregulated genes were _LOC105370794_, _DDR2_, _TMEM200A_, _GIMD1_, _NMNAT3_, _NCKAP5_, and _LOC102723862_, whereas _TPH1_, _LOC107985155_, and _LTBP1_, were \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline
**Library Name** & **Total Reads** & **Total Read (bp)** & **GC (\%)** & **AT (\%)** & **Q20 (\%)** & **Q30 (\%)** \\ \hline DW-1 & 42,923,432 & 4,335,266,632 & 50.81 & 49.19 & 98.04 & 94.69 \\ DW-2 & 35,861,160 & 3,621,977,160 & 50.51 & 49.49 & 98.33 & 95.15 \\ DW-3 & 36,625,918 & 3,699,217,718 & 51.06 & 48.94 & 98.22 & 95.07 \\ GP-P1 & 40,523,746 & 4,092,898,346 & 51.19 & 48.81 & 98.08 & 94.72 \\ GP-P2 & 38,146,816 & 3,852,828,416 & 50.72 & 49.28 & 98.28 & 95.11 \\ GP-P3 & 49,818,356 & 5,031,653,956 & 50.98 & 49.02 & 98.29 & 95.16 \\ GP\_C1 & 35,192,798 & 3,554,472,598 & 51 & 49 & 98.24 & 95.06 \\ GP\_C2 & 33,870,946 & 3,420,965,546 & 50.7 & 49.3 & 98.15 & 94.85 \\ GP\_C3 & 44,721,472 & 4,516,868,672 & 50.82 & 49.18 & 98.23 & 94.98 \\ GP\_CR1 & 34,605,546 & 3,495,160,146 & 50.48 & 49.52 & 98.05 & 94.63 \\ GP\_CR2 & 42,126,968 & 4,254,823,768 & 50.9 & 49.1 & 98.08 & 94.89 \\ GP\_CR3 & 32,703,588 & 3,303,062,388 & 50.73 & 49.27 & 98.22 & 95.02 \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Summary of the raw data obtained by RNA sequencing. Figure 2: Proportion of mapped reads on the human reference genome in each library. significantly upregulated. Moreover, four lncRNAs (_LINC00498_, _LINC00539_, _LINC01359_, and _LINC02355_) were found to be differentially expressed following GP-C treatment. Figure 4: Volcano plot displaying the top ten differentially-expressed genes in each condition. Differentially-expressed genes were identified in four different groups such as GP-P (**A**), GP-C (**B**), GP-CR (**C**), and GP-All (**D**) as compared to the control group. Green and red colors indicate down- and upregulated genes, respectively. The top ten ranking genes are indicated by respective gene names based on the Manhattan distance using the VolcaNoseR program. Figure 3: Number of DEGs identified in each condition. Green and red colors indicate down- and upregulated genes, respectively. Four comparisons were conducted: GP-P (GP extract from the plant leaves), GP-C (GP extract from the callus), GP-CR (GP extract from the callus treated with radiofrequency), and GP-all (Combination of all three GP extracts). GP, _Gynostemma pentaphyllum_; DEGs, differentially-expressed genes. Of the 90 DEGs in the GP-CR treatment (Table S3), five genes, _LOC107984359, TMEM26, LOC105370794, LOC105376934_, and _NMUR2_, were identified as significantly downregulated, whereas three genes, _LOC107985495, LTBP1_, and _CAMKMT_, were identified as significantly upregulated (Figure 4C). Further, we identified four lncRNAs, _LINC00539, LINC00877_, _LINC02224_, and _LINC02355_, under GP-CR treatment (Table S3). Under GP-all analysis, 176 DEGs were identified (Table S4). The most significantly upregulated genes were _SLC26A11_, _LTBP1_, _LOC105377548, TPH1_, and _CFAP70_, whereas three genes, _CRHR2_, _LOC107986720_, and _LOC105377128_, were significantly downregulated (Figure 4D).