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Immunofluorescence, confocal microscopy image acquisition and colocalization analysis For the immunofluorescence analysis of endogenous proteins, RAW 264.7 murine macrophages (ATCC, TIB-71) were seeded on untreated coverglass using a four-chamber 35-mm dish with 20-mm microwells (Cellvis, D35C4-20-1.5-N), incubated at 37 C for 30 min, then fixed with 3.7% paraformaldehyde for 8 min, permeabilized with 0.01% saponin and blocked with 1% BSA in PBS

While the phyB mutant has a long hypocotyl at both 20C and 27C, due to constitutively induced thermomorphogenesis, the pif4 mutant fails to induce hypocotyl elongation at 27C due to its impaired thermomorphogenesis pathway (Figure 5E)

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The cultured SGN explants from WT or gpx1 / mice were treated with peroxynitrite and 2 mM NAC for 48 h (Figures 6A, 7A)