## 3 Results
### Effect of Electroporation on SERCA mRNA and Protein Expression  
The electroporation of cells can have several negative effects on homeostasis (Figure 1B). The disruption of ion balance and ATP loss due to electroporation have severe effects on the normal function of ion pumps. It was reported that low expression levels of SERCAs and  
\begin{table}
\begin{tabular}{c c c c c} \hline \hline
**Antibody** & **Host** & **Type** & **Dilution** & **Supplier** & **Cat. No.** \\ \hline NKA\(\alpha\)1 Tyr10 & Rabbit & Monoclonal & 1:500 & CST & 13566 \\ NKA\(\beta\)1 & Rabbit & Monoclonal & 1:100 & CST & 62849 \\ NKA\(\beta\)2 & Rabbit & Polyclonal & 1:500 & TFS & PA5-121248 \\ NKA\(\beta\)3 (C\(\Omega\)298) & Rabbit & Monoclonal & 1:1000 & Invitrogen & MA5-37934 \\ SERCA1 & Rabbit & Polyclonal & 1:1000 & CST & 4219 \\ SERCA2 & Rabbit & Monoclonal & 1:1000 & CST & 9580 \\ Pln & Rabbit & Monoclonal & 1:500 & CST & 14562 \\ Pln Serife/Thr17 & Rabbit & Polyclonal & 1:1000 & CST & 8496 \\ ACC & Rabbit & Monoclonal & 1:1000 & CST & 3676 \\ AMPK\(\alpha\) & Rabbit & Polyclonal & 1:1000 & CST & 2532 \\ pAMPK\(\alpha\) Thr172 & Rabbit & Monoclonal & 1:1000 & CST & 2535 \\ pACC Ser79 & Rabbit & Polyclonal & 1:1000 & CST & 3661 \\ NKA\(\alpha\)1 & Mouse & Monoclonal & 1:2000 & Merck & 05-369 \\ NKA\(\alpha\)2 & Rabbit & Polyclonal & 1:4000 & Merck & AB9094-I \\ NKA\(\alpha\)3 & Mouse & Monoclonal & 1:1000 & TFS & MA3-915 \\ PXYD1 & Rabbit & Polyclonal & 1:1000 & Invitrogen & PA5-79288 \\ pFXYD1 Ser68 & Rabbit & Polyclonal & 1:500–2000 & TFS & PA5-99971 \\ \hline \hline \end{tabular} \(\mathrm{CST}\): Cell Signaling Technology; TPS: Thermo Fisher Scientific.  
\end{table}
Table 1: List of primary antibodies used in this study.  
PMCAs may play a role in the higher sensitivity of tumor cells compared to normal cells to calcium electroporation [30; 31; 49]. Thus, we investigated how electroporation may affect the expression of SERCA isoforms 1, 2, and 3 and the expression and phosphorylation of the SERCA regulator phospholamban (PIn).  
Regarding mRNA levels, a significant decrease in the expression of SERCA1 (_Atp2a1_) was observed within the first 24 h following electroporation, with both monopolar and H-FIRE pulses (Figure 4A,D). The same was also observed at the protein level, where both monopolar and H-FIRE pulses caused a downregulation in expression at 4 and 24 h (Figure 4G,J). On the contrary, the mRNA expression of SERCA2 (_Atp2a2_) remained stable, as can be seen in Figure 4B,E, but we detected a downregulation in the protein level, which was significant at 4 h for H-FIRE pulses but not for monopolar pulses (Figure 4H,K). SERCA3 mRNA levels were very low (Figure 2), but we nevertheless detected an increase in the expression 4 h after electroporation with both monopolar and H-FIRE pulses (Figure 4C,F). We also evaluated the mRNA expression of phospholamban (_PIn_) but were unsuccessful in detecting it at the protein level. At the mRNA level, phospholamban levels were significantly increased at the basal level when comparing 4 to 24 h (Figure 4I,L), but 24 h after electroporation with higher electric fields, the expression was reduced to the 4 h basal levels.