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Fig. 4. Expression of CBP in late passage HMEC-E6 cells promotes CBP binding to the LAMA3A promoter and increases LAMA3A promoter activity and laminin-5 expression. (A) CBP protein expression in (1) early passage HMEC-E6 cells (passage 10), (2) late passage HMEC-E6 cells (passage 18; ${Phi}$ ), (3) late passage HMEC-E6-CBP^– vector controls (passage 18; (–)CBP), and (4) late passage HMEC-E6-CBP⁺ cells (passage 18; (+)CBP). Equal amounts of protein lysate were added per lane. Actin was used as a loading control. (B) Exogenous expression of CBP in late passage HMEC-E6-CBP⁺ cells promotes CBP occupancy of the 277 bp AP-1-rich region of the LAMA3A promoter. ChIP was performed in (1) early passage HMEC-E6 cells, (2) late passage HMEC-E6 cells (passage 17; ${Phi}$ ) and (3) late passage HMEC-E6-CBP^– vector controls (passage 17; CBP(–)) and compared with late passage HMEC-E6-CBP⁺ cells (passage 17; CBP(+)). Input controls test the integrity of the DNA samples. (C) LAMA3A promoter activity was measured in (1) early passage HMEC-E6 cells (E6E; passage 10), (2) late passage HMEC-E6 cells (E6L no tx; passage 18), and (3) late passage HMEC-E6-CBP^– vector controls (E6L(–)CBP; passage 18) and compared with late passage HMEC-E6-CBP⁺ cells (E6L(+)CBP; passage 18). Data represent two independent experiments performed in triplicate. (D) Laminin-5 protein expression in (1) early passage HMEC-E6 cells (passage 10), (2) late passage HMEC-E6 cells (passage 18; ${Phi}$ ), (3) late passage HMEC-E6-CBP^– vector controls (passage 18; (–)CBP), and (4) late passage HMEC-E6-CBP⁺ cells (passage 18; (+)CBP). Equal amounts of protein lysate were added per lane. Actin was used as a loading control.

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