Figure 4
Figure 4. Validation of KLF1 and LMO2 as c-myb direct target genes. (A) KLF1 promoter-specific PCR performed in a representative ChIP experiment. CTSG promoter-specific PCR is shown as a positive control for immunoprecipitation. ChIP and PCRs were performed in triplicate. (B) c-myb transactivation of KLF1 promoter-driven luciferase expression. The amounts (in micrograms) of cotransfected plasmids are reported. In the panel below, protein extracts from the transfected cells were tested for c-myb expression levels by Western blot. Luciferase levels (mean ± 2 SEM;n = 3) are normalized by setting the empty pXP1/pCMV6XL5 vectors-transfected sample as = 1. Error bars represent SD. *P ≤ .05 versus pXP1KLF1-685+empty pCMV6XL5-transfected sample. (C) LMO2 promoter-specific PCR performed in a representative ChIP experiment. CTSG promoter-specific PCR is reported as a positive control for immunoprecipitation. ChIP and PCRs were performed in triplicate. (D) c-myb transactivation of LMO2 promoter-driven luciferase expression. The amounts (in micrograms) of cotransfected plasmids are reported. In the panel below, c-myb expression levels were detected by Western blot in protein extracts from the transfected cells. Luciferase levels (mean ± 2 SEM; n = 3) are normalized by setting the empty pXP1/pCMV6XL5 vectors-transfected sample as = 1. Error bars represent SD. *P ≤ .05 and **P ≤ .01 versus pXP1LMO2-175+empty pCMV6XL5-transfected sample. n = number of experiments.

Validation of KLF1 and LMO2 as c-myb direct target genes. (A) KLF1 promoter-specific PCR performed in a representative ChIP experiment. CTSG promoter-specific PCR is shown as a positive control for immunoprecipitation. ChIP and PCRs were performed in triplicate. (B) c-myb transactivation of KLF1 promoter-driven luciferase expression. The amounts (in micrograms) of cotransfected plasmids are reported. In the panel below, protein extracts from the transfected cells were tested for c-myb expression levels by Western blot. Luciferase levels (mean ± 2 SEM;n = 3) are normalized by setting the empty pXP1/pCMV6XL5 vectors-transfected sample as = 1. Error bars represent SD. *P ≤ .05 versus pXP1KLF1-685+empty pCMV6XL5-transfected sample. (C) LMO2 promoter-specific PCR performed in a representative ChIP experiment. CTSG promoter-specific PCR is reported as a positive control for immunoprecipitation. ChIP and PCRs were performed in triplicate. (D) c-myb transactivation of LMO2 promoter-driven luciferase expression. The amounts (in micrograms) of cotransfected plasmids are reported. In the panel below, c-myb expression levels were detected by Western blot in protein extracts from the transfected cells. Luciferase levels (mean ± 2 SEM; n = 3) are normalized by setting the empty pXP1/pCMV6XL5 vectors-transfected sample as = 1. Error bars represent SD. *P ≤ .05 and **P ≤ .01 versus pXP1LMO2-175+empty pCMV6XL5-transfected sample. n = number of experiments.

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