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In HeLa cells, siRNA transfection was carried out through reverse transfection protocol with HiPerFect (QIAGEN). Six microliters of 10 μM siRNAs targeting human siKLC1 or siKLC2 was mixed with 12 μl of HiPerFect in high-glucose DMEM (Sigma-Aldrich) and combined with 1 × 105 cells per well of a six-well plate. Samples were collected 72 hours after the transfection. For rescue experiments, corresponding mouse KLC1/2 cDNAs (near identical proteins) were transiently transfected so that the siRNA would not recognize the targeting sequence.
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