Figure 6.

CsFnl7.1 can interact with CsDLP6 and CsGLP1. (a) Yeast two‐hybrid analysis. CsFnl7.1 protein was fused to the DNA‐binding domain (BD) to generate the prey construct. CsDLP6 and CsGLP1 proteins were fused to the GAL4 activation domain (AD) to generate the bait constructs. DDO, double drop‐out medium (SD/‐Trp‐Leu) containing X‐α‐gal; QDO, quadruple drop‐out medium (SD/‐Trp‐Leu‐His‐Ade) containing X‐α‐gal. BD‐lam is a negative control that encodes a fusion of human lamin C protein, and the GAL4 DNA‐BD. BD‐53 and AD‐T are positive control that encode a fusion of murine p53 protein and the GAL4 DNA‐BD or a fusion of SV40 large T antigen and the GAL4 DNA‐AD. (b) In vitro pull‐down analysis using anti‐HIS and anti‐GST antibodies. The left lanes are control groups using GST and HIS‐CsDLP6 or HIS‐CsGLP1 as inputs. The right lanes are test groups using GST‐CsFnl7.1 and HIS‐CsDLP6 or HIS‐CsGLP1 as inputs. Using anti‐HIS Abs, the presence of a band indicates that the pulled down proteins contain HIS‐CsDLP6 or HIS‐CsGLP1 in the test group, suggesting that the HIS‐fusioned proteins could be pulled down by GST‐CsFnl7.1 but not by GST only. Using anti‐GST Abs, the presence of bands further verified the interactions. (c) Quantitative PCR expression analysis of CsDRP6 and CsGLP1 in necks of Jin5‐508 and YN. (d) Confirmation of CsDRP6 and CsGLP1 expression levels in the necks of transgenic cucumber lines at 0 day post‐pollination as assessed by quantitative PCR. WT: wild type (D8). OE4, OE7 and OE8 are the three CsFnl7.1 overexpression lines. Each value denotes the mean relative expression level of three replicates. The short lines show the means of three biological replicates.