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. 2019 Mar 28;9:65. doi: 10.3389/fcimb.2019.00065

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Copyright © 2019 Cao, Lyu, Jia, Wang, Du, Pan, Li, Xing, Xiao, Ma and Zhang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The NRF1 and SMAD2 interactors identified on a Mtb proteome microarray and validation by His pull-down assay. (A) Silver staining to detect the purified NRF1-myc and SMAD2-myc. (B) NRF1 and SMAD2 binding study with Mtb proteome microarray. Each array contained Cy3 labeled BSA as a positive control (marked with red arrows) and BSA as a negative control (marked with blue arrows). Positive proteins were marked with yellow arrows. The cutoff for calling positive candidates was set as signal-to-noise (SNR) ratio ≥ 1.4 and Z-score ≥ 3.0. (C) His tagged -Rv0577 pull-down assay with endogenously expressed NRF1. (D) His tagged -Rv0577, -Rv3153, -Rv2117, and -Rv2423 with endogenously expressed SMAD2, respectively. Protein separation and detection were performed using an automated capillary electrophoresis system. The results are representative of two independent experiments.