(A) Transcripts for rCBF-A and FSP1 increase in PCR cycles after rCBF-A induction. (B) ChIP assay shows formation of the FTS-1 complex in epithelial cells expressing rCBF-A (+Dox; doxycycline) compared to 3T3 cells. IgG was a loading control. Immunoprecipitated DNA was used in PCR with primers for FTS-1. (C) Nuclear protein from MCT/rCBF-A⁺ or 3T3 fibroblasts form FTS-1 complexes in EMSA with similar affinity, challenged by 10× or 20× molar excess of nonlabeled FTS-1 probe (Cold probe). MCT cells were the negative control. (D) Differential expression of epithelial and mesenchymal protein markers in MCT/rCBF-A⁺ cells at different hours after induction of rCBF-A compared with 3T3 fibroblasts. Coo, Coomassie staining. (E) Increased expression of CBF-A and FSP1 in UUO undergoing EMT (28% and 26%, respectively) as measured by qRT-PCR and represented graphically in arbitrary units. Included for comparison are E-cadherin, α-SMA, and KAP-1. (F) CBF-A (yellow/orange) accumulates in the nuclei of tubular and interstitial cells from the UUO kidney in FSP1.GFP mice. FSP1⁺ fibroblasts expressing GFP under the control of the FSP1 promoter were recolored dark blue, and nuclei were counterstained red. Normal contralateral kidney tissue demonstrates CBF-A in the tubular cytoplasm (speckled green). In the cytoplasm, CBF-A and FSP1 staining merge to become light blue. Original magnification, ×630.