Extended Data Fig. 2: Dow1 encodes a BAHD acyltransferase protein localized in the cytosol and membrane.
From: A sucrose ferulate cycle linchpin for feruloylation of arabinoxylans in plant commelinids
a, Gene structure of Dow1 and locations of the Mu insertions in two independent alleles. Exons are closed boxes, and introns are lines. b, Co-transfecting the maize protoplasts, Arabidopsis protoplasts, and tobacco leaves with GFP-fused DOW1 and RFP-fused BiP, GFP-fused DOW1 and RFP-fused BiP, RFP-fused DOW1 and GFP-fused BiP. Scale bar=5 μm for maize protoplasts, 5 μm for Arabidopsis protoplasts, and 20 μm for tobacco leaves. BiP-RFP was used to indicate endoplasmic reticulum (ER). BF, bright field. c, Immunoblot analysis of total proteins and nucleus proteins. DOW1 does not accumulate in the nucleus. Anti-H3 was used to indicate the nucleus. T: total proteins from 3-day-old seedlings; N: nuclear protein. d, Immunoblot analysis of cytoplasm non-membranous proteins and cell total membranous proteins. Cyt: Cytoplasm non-membranous proteins; CM1: cell total membranous proteins; CM2: 5 times the concentration of CM1; W: supernatant of washing cell total membranous proteins; Anti-BiP was used to indicate ER. Abs, antibodies. e, Immunoblot analysis of the fractions obtained from maize seedlings by a sucrose density gradient centrifugation. The distribution pattern of DOW1 is identical to that of BiP, which is an ER marker, indicating that DOW1 is localized to ER. Anti-BiP and anti-PIP1s antibodies were used to indicate ER and plasma membrane. Abs, antibodies. f, RT-qPCR analysis of the Dow1 transcript levels in major maize tissues. Error bars=mean ± SD (n = 3 replicates of assays with independent samples). Total RNA was isolated from all tissues, reverse-transcribed, and used as templates. Kernels are indicated as DAP (days after pollination). Experiments were repeated independently with similar results at least 3 times (b-e).
