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After rinsing, slides were incubated with the secondary antibody Alexa Fluor 488 goat anti-rabbit (A-11008 Invitrogen - Thermo Fisher Scientific) or Alexa Fluor 488 donkey anti-goat (A-11055 Invitrogen - Thermo Fisher Scientific) 1:500 in PBS 1 0.5% BSA 0.1% saponin (w/v/v) for 2 h at room temperature