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Figure 3 | BMC Plant Biology

Figure 3

From: Involvement of co-repressor LUH and the adapter proteins SLK1 and SLK2 in the regulation of abiotic stress response genes in Arabidopsis

Figure 3

LUH has repressor activity and SLK1, SLK2 acts as adapter protein for LUH recruitment. (A) Schemes of the reporter and effector constructs. In the reporter plasmid 5X Gal4 binding site were fused to CaMV 35S promoter and LUC gene. Renilla luciferase (R-LUC) reporter is used as an internal control for transfection. In the effector plasmid, SLK1, SLK2 and LUH gene were fused to the Gal4-BD under the control of CaMV 35S promoter. For effector LUH and LUFS without Gal4-BD, the gene was fused to CaMV 35S promoter and noplaine synthase terminator. (B) Arabidopsis protoplasts were transfected with luciferase reporter construct (10 microgram DNA) plus Renilla luciferase (0.5 microgram DNA) together with the effector construct. The ratios of LUC/R-LUC indicate relative reporter activity. Twenty microgram of pXSN (vector), Gal-BD, SLK1-BD, SLK2-BD and LUH-BD DNA was used for 2X and ten microgram for 1X in repression assay. (C) To determine adapter role of SLK1 and SLK2, the Arabidopsis protoplasts were transfected with reporters as above together with ten microgram of each Gal-BD, SLK1-BD, SLK2-BD DNA and 20 microgram of CaMV 35S::LUH or CaMV 35S::LUFS DNA. The ratios of LUC/R-LUC indicate relative reporter activity. 20 μM TSA was used to inhibit HDAC activity in protoplasts. Error bars are SE (n = 3). Asterisks indicate values that are significantly different from the pXSN vector control for (B) and Gal-BD for (C) (*P <0.05, Student’s t test).

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