Fig. 5

CaCrz1 binds in vitro and in vivo to its own promoter. (a) Locations of three predicated CaCrz1-binding motifs (boxed and within Probe 1, Probe 2 and Probe 3 sequences) based on the consensus motif we discovered in this study and one predicated CaCrz1-binding motif (boxed and within the Probe 4 sequence). Locations of the ChIP PCR primer pair [CHIP_CRZ1_F和CHIP_CRZ1_R] are indicated with broken lines above and under their corresponding sequences, respectively. (b) DIG-labelled Probe 1_EMSA_CRZ1_F/R was added into samples in Lanes 1–3. DIG-labelled Probe 2_EMSA_CRZ1_F/R was added into samples in Lanes 4–6. DIG-labelled Probe 3_EMSA_CRZ1_ F/R was added into samples in Lanes 7–9, and DIG-labelled Probe 4_EMSA_CRZ1_ F/R was added into samples in Lanes 10–12. Unlabelled probes 1, 2, 3 and 4 were added into samples in Lanes 3, 6, 9 and 12, respectively. Purified His6-Crz1 protein of 1 μg was added into Lanes 2, 3, 5, 6, 8, 9, 11 and 12. (C) Detection of CaCrz1 binding to its own promoter in vivo by ChIP analysis. The same pair of strains were treated and their whole cell extracts were immunoprecipitated as in Fig. 3c. PCR reactions were carried out with ChIP primers CHIP_CRZ1_F和CHIP_CRZ1_R. The lower panel is the inverse image of the topper panel, which is for a better view of the PCR band in the second lane