It is worth to mention here that the cell transfection efficiency in these experiments is about 35 40%, as determined by green fluorescence protein transfection control, and it is in accordance with the bibliography [37], [38].
It is noteworthy to say among three mentioned constructs 2RMPG-2H1 showed the highest transfection efficiency.
Despite the advantages mentioned, low immunogenicity and expression levels, low transfection efficiency and targeting specificity remain the main obstacles compared to conventional vaccines [ 6, 7].
Note that transfection efficiencies routinely reached 70 90%.
Transfection efficiency has been calculated by counting GFP positive cells in the above mentioned flow cytometer using Guava Express plus software (v3.6.1).
As mentioned above in response to Reviewer 1, it is not common to transfect lymphoblastoid cells to target a putative causal gene in CNV patients due to low transfection efficiency and survival of lymphoblastoid cells upon transfection.
Transfection efficiency was determined at 24 h post-transfection.
Transfection efficiency was evaluated by RT-qPCR.
(J) Id3 siRNA transfection efficiency was found to be ~75%.
Cells were sorted for GFP 3 days post-transfection if transfection efficiency was below 90% (based on visual inspection).
