For the MTT assay, approximately 5 × 104 cells were seeded on a test bed Si NN-patch in a 24-well plate. At each measurement point, 200 μl of MTT solution was added to the wells and incubated for 4 hours. The cell medium was excavated, and 400 μl of dimethyl sulfoxide (Sigma-Aldrich, USA) was added to dissolve precipitated formazan. Approximately 100 ml of the solution was transferred to a 96-well plate and measured with a microplate reader (SpectraMax Plus 384 reader, Molecular Devices, USA) at 570 nm. For the LDH assay, the cells were evaluated at 24 and 48 hours after treatment of 0.25% (v/v) Triton X-100 (Sigma-Aldrich, USA) as a positive control. After incubation, 100 ml of supernatant from each well was removed and centrifuged at 1000 rpm for 5 min. Ten microliters of aliquot was transferred to additional microplate, and 100 μl of LDH assay buffer mix was added to the wells and incubated for 30 min. The absorbance was measured with a microplate reader (SpectraMax Plus 384 reader, Molecular Devices, USA) at 450 nm. For the confocal microscopy analysis, the cells were fixed with 4% (v/v) paraformaldehyde (Sigma-Aldrich, USA) in PBS for 15 min, stained with 4′,6-diamidino-2-phenylindole (500 nM, Fisher Scientific, USA) for 2 min, and then mounted with an antifade reagent (Gold Antifade Mountant, Invitrogen, USA). For the flow cytometry (FACS) analysis, the cells injected with GAPDH Cy3-siRNAs were trypsinized, washed several times with PBS, and then fixed in 0.5% (v/v) paraformaldehyde for 1 hour. To confirm the expression of GAPDH, approximately 3 × 104 cells were seeded on a 12-well plate and incubated for 24 hours. The efficacy in silencing GAPDH was evaluated by analyzing fluorescent intensity at 450 nm with a standard GAPDH assay kit (Abcam, USA).

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