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. 2021 Jun 18;254(1):8. doi: 10.1007/s00425-021-03657-6

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Fig. 6 — OsJAZ11 interacts with OsSPX1 to regulate Pi homeostasis. a Yeast two-hybrid assay showing interaction of OsJAZ11 (pBD-OsJAZ11) with OsSPX1 (pAD-OsSPX1). Serial dilutions of Y2H Gold strain co-transformed with AD (prey) and BD (bait) clones were spotted on DDO (SD-Leu/-Trp), TDO (SD- Leu/-Trp/-His), QDO (SD- Leu/-Trp/-His/-Ade) and QDO/A/X (QDO + Aureobasidin A + X-α-Gal). b GST pull-down assay showing interaction of OsJAZ11-GST and OsSPX1-6XHis. GST (negative control) and OsJAZ11-GST were immobilized on glutathione-agarose beads and incubated with OsSPX1-6XHIS (input protein). Pulled-down complexes were detected by immunoblotting using anti-GST and anti-HIS antibodies. c EMSA assay showing no effect of OsJAZ11 on OsSPX1-mediated inhibition of OsPHR2 binding to P1BS. OsPHR2-6XHIS (0.7 μg) and 50 ng DIG-labeled P1BS probe was incubated with either 7.5 μg SPX-6HIS (lane 3), SPX-6XHIS + 7.5 μg OsJAZ11-GST (lane 4) or SPX-6XHIS + 7.5 μg GST (lane 5). d–g Relative expression levels of OsSPX1, OsSPX2, OsSPX3 and OsSPX5 in roots of 30-day-old WT and OsJAZ11 transgenics. Each bar is the mean of three independent replicates with standard error. Different letters on top of each bar denote significant differences determined by one-way ANOVA followed by Duncan’s multiple comparison test (α < 0.05) (n = 3)