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Experiments were conducted in quadruplicate

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For IF, AT2 cells were fixed with 4% paraformaldehyde at room temperature for 15 min, incubated with 0.1% Triton X-100 for 5 min to permeate the cell membrane, and blocked with 5% goat serum at room temperature for 30 min, followed by primary antibody incubation (1:100) overnight at 4C

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Consistent with these associations of IGF1 and IGFBP5 mRNA, there was significantly higher CD274 expression in Firehose LegacyPCatumors in the highest vs