2 4 amidinophenyl 1h indole 6 carboxamidine dapi

Peritoneal macrophages (1.0 × 10⁶ cells/mL) were seeded in 0.3 mL of medium in 24-well plates. The cells were treated with isocorydine (52.03 µM) for 2 h both in the presence and absence of LPS (50 ng/mL). The cells were then processed for further analyses. The cells on coverslips were fixed using 4% paraformaldehyde for 20 min and permeabilized using 0.3% Triton X-100 (Solarbio) in PBS for 20 min. Then, the cells on the coverslips were blocked for 1 h at 37°C in PBS containing 3% bovine serum albumin (BSA) and incubated with primary antibodies recognizing NFκB p65 (CST, #8242) at 4°C overnight. Next day, the cells were incubated with FITC Goat anti-Rabbit IgG (BA1105, Boster, Wuhan, China) and counterstained with 2-(4-amidinophenyl)-1H-indole-6-carboxamidine (DAPI) (Solarbio). Images were captured using a Zeiss LSM900 confocal microscope (Zeiss, Oberkochen, Germany) with a Plan-Apo Chromat 63×/1.40 oil objective.