40S Ribosome Biogenesis Co-Factors Are Essential for Gametophyte and Embryo Development
Figure 5
Analysis of T-DNA insertion mutants.
A, Positions of the T-DNA within the genes are shown. Accession numbers of the plant lines and the name used here is given. The base position verified by T-DNA mapping (Table S1) is indicated on the left or right border of the insertion. Black arrows indicate primer binding sites used for the analysis (Table S2). B, Segregation state of insertion lines was verified with PCR. The T-DNA left border primer was combined either with the forward (lane 1) or reverse genomic primer (lane 2). For lane 3 the forward and reverse genomic primers were used. C, mRNA-levels in wild-type and mutants were analyzed by qRT-PCR. Values were normalized to ACT2 and the wild-type level was set to 100% for comparison to the expression level in mutants. Oligonucleotides are listed in Table S2. D, Protein levels of atNob1 in WT and nob1+/− were determined by immunodecoration of plant extract with αNOB1 or αACT2 antibodies (loading control). E, Protein levels of atEnp1 in WT and enp1+/− were determined by immunodecoration of plant extract with αEnp1 or αAct2 antibodies (loading control).