Fig. 9

Immunoprecipitation (Co-IP) of a TF complex and phosphorylation of BbOsrR2 and BbOsrR3. A Co-IP assays using tagged versions of BbOsrR1, BbOsrR2, and BbOsrR3 as detailed in the “Methods” section. B Co-IP assays using tagged versions of BbOsrR1 and BbClp1. C Co-IP assays using tagged versions of BbOsrR2, BbOsrR3, and BbClp1. D Co-IP assays using tagged versions BbOsrR2 and BbOsrR3 in the ΔBbOsrR1 strain. E Co-IP assays using tagged versions of BbOsrR3 and BbClp1 in the ΔBbOsrR2 strain. F Co-IP assays using tagged versions of BbOsrR2 and BbClp1 in the ΔBbOsrR3 strain. G Co-IP assays of tagged versions of BbOsrR2 and BbOsrR3 in the ΔBbClp1 strain. Anti-β-tubulin antibody was used as an internal standard. H Assays for phosphorylation of BbOsrR2 and BbOsrR3 in the wild type and ΔBbClp1 strains. Phosphorylation of BbOsrR2 and BbOsrR3 from fungal cultures was performed using the Myc-tagged version of BbOsrR2 and the His-tagged version of BbOsrR3 in cells grown under normal and oxidative stress for 30 min after Western blotting via Phos-tag™ SDS-PAGE as described in the “Methods” section. BbOsrR2 and BbOsrR3 proteins were blotted with their respective (anti-)tag antibodies. The relative amounts of the phosphorylated BbOsrR2 and BbOsrR3 as compared to their total protein levels for each sample were measured by densitometric analysis of bands using the ImageJ software