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After stimulations, chamber slides were fixed with 4% paraformaldehyde for 20 minutes, blocked in 10% goat serum in phosphate-buffered saline (PBS) with 0.1% saponin for 1 h and then stained with integrin-1 antibody (M-106, Santa Cruz Biotechnology, Santa Cruz, CA, USA) in 0.1% saponin in PBS for 2 h at room temperature