Figure 6.

Pti4 Is Required for Binding of SEBF to the PR-10a Promoter.

(A) Pti4 transcript is less abundant in Pti4 VIGS lines. The bar diagram illustrates the abundance of Pti4 transcript in PVX-Pti4 VIGS lines (VIGS#1 to VIGS#4) relative to PVX-00 lines (PVX-00#1 to PVX-004). Data for each of the four biological replicates (each bar) are averages of three technical replicates, and errors are equal to 1 sd.

(B) Reduced Pti4 prevents SEBF binding to the PR-10a promoter. ChIP results were analyzed by qPCR. The amounts of the different transcription factors binding to PR-10a were relative to their recruitment to the Actin gene PoAc97 (see Methods). ChIPs from two independent PVX-Pti4 VIGS lines (VIGS#1 and VIGS#2) and from a VIGS line containing the empty pGR106 vector (PVX-00) were performed with uninduced tissues. ChIP was conducted with an anti-SEBF (SEBF) or an anti-Pti4 (Pti4) antibody. Data for each bar are from two biological replicates, and errors are equal to 1 sd.

(C) SEBF is still present in the Pti4 VIGS lines. The two top panels are immunoblot analysis of SEBF and Why1 proteins extracted from the PVX-00 line and from the PVX-Pti4 VIGS lines 1 and 2 (VIGS#1 and VIGS#2). An anti-SEBF or anti-St Why1 antibody was used. The bottom panel is a Ponceau staining of the membrane, shown in the top panels, as a loading control.

(D) PR-10a transcripts are more abundant in the Pti4 VIGS lines. The bar graph illustrates the abundance of PR-10a transcript present in the PVX-00 lines (PVX-00#1 to PVX-004) and in the PVX-Pti4 VIGS lines (VIGS#1 to VIGS#4). Data for each of the four biological replicates (each bar) are averages of three technical replicates, and errors are equal to 1 sd.