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Then, the cells were fixed with 4% paraformaldehyde, permeabilized with 0.02% Triton-X-100/1 × phosphate-buffered saline (PBS) and blocked in a blocking fluid (1 × PBS + 10% fetal bovine serum (FBS) and 1% bovine serum albumin (BSA)) (Thermo Fisher Scientific, Waltham, USA)