Fig 1: Hypoxia–reoxygenation (H-R) challenge increases the abundance of miR-210 promoter-bound transcriptional coactivators, p300 and CBP, concomitant with a decrease in abundance of the miR-210 promoter-bound transcriptional corepressors, NCoR1 and SMRT, through NF-?B activation. (A–D) Quantitative ELISA determining the relative abundance of p300 (A), CBP (B), NCoR1 (C), and SMRT (D) in the lysates emanating from the reverse crosslinked miR-210 promoter pull-down fragments from H-R challenge-subjected cells ectopically expressing the da-I?Ba mutant. The relative abundance of ß-actin (Supplementary Figure S4A) and chromatin-associated TBP (Supplementary Figure S4B) in the native cellular lysates was determined using sandwich ELISA immunoassays to validate equitable inputs across the experimental lysates subjected to miR-210 promoter pull-down. The relative abundance of chromatin-associated histone H3 (Supplementary Figure S4C) was determined in the reverse crosslinked miR-210 promoter pull-down fragment to validate that equitable fraction of the miR-210 promoter was pull-down across the experimental lysates. The ectopic expression of the HA-tagged da-I?Ba mutant was validated using sandwich ELISA immunoassay performed against the HA tag (Supplementary Figure S4D). NF-?B transcriptional activity (Supplementary Figure S4D, bottom panel) was determined in the native lysates to corroborate and validate the translative effects of the ectopic expression of the da-I?Ba mutant. Data are expressed as experimental blank-corrected absorbances (O.D) measured at ?450 (450 nm) normalized to fold change values. All data are expressed as mean fold change ± S.D. values from three technical replicates for each of the four biological replicates belonging to each experimental group (n = 4). * p = 0.05; *** p = 0.001; **** p = 0.0001; ns: not significant (p > 0.05). S.D.: standard deviation.
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