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. 2018 Mar 16;9:267. doi: 10.3389/fpls.2018.00267

Figure 4.

Figure 4

Root nodule symbiosis (RNS) related gene expression is promoted by IPD3 and phosphoablative/-mimetic IPD3 variants. (A) The relative expression levels of RNS-specific genes revealed by real-time PCR using the transgenic hairy roots of M. truncatula at 4 wpi. These experiments were repeated three biological times. More than ten plants were used for RNA extraction. (B) Transactivation assay of IPD3 in Nicotiana benthamiana leaves. 3xHA-IPD3-WT, 3xHA-IPD3-2A, 3xHA-IPD3-2D, 3xHA-IPD3-8A, or 3xHA-IPD3-8D was co-injected in N. benthamiana leaves with the pERN1:GUS reporter. GUS activity of leaf discs was determined histochemically and quantitatively as described previously (Boisson-Dernier et al., 2005). (C) Western blots revealed the protein content of 3xHA-IPD3-WT, 3xHA-IPD3-2A, 3xHA-IPD3-2D, 3xHA-IPD3-8A, 3xHA-IPD3-8D in N. benthamiana leaves. These experiments were repeated three times. The error bars indicate standard error (n = 3). The asterisk indicates a significant decrease relative to the control (EV) with Student's t-test (*P ≤ 0.01).