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Fig. 2 | BMC Molecular Biology

Fig. 2

From: Molecular analysis of NPAS3 functional domains and variants

Fig. 2

The bHLH and PAS domains are required for high-affinity interaction of NPAS3 and ARNT. Domain constructs used indicated in Fig. 1a. Western blots of HaloTag pull-down of HA-tagged NPAS3 domains by HaloTag-ARNT. Blots include lanes for input sample, unbound sample taken after binding reaction, and protein eluted off of resin after washing. Bands in the top panel of pull-down lane indicate physical association of HA-tagged NPAS3 domain construct with the HaloTag-ARNT construct. Middle and bottom panels are results for the HaloTag-ARNT and HaloTag-empty constructs. As HaloTag constructs are covalently linked to the resin, they will be depleted in the pull-down sample, demonstrating functionality of the HaloTag. Molecular weight of HaloTag-ARNT, 121 kDa, HaloTag construct with no cDNA: 34 kDa. Blot legend with expected sizes: a bHLH–PAS, 49.4 kDa, b PAS-TAD, 98.2 kDa, c bHLH, 13 kDa, d PAS, 35.8 kDa, e TAD, 53.2 kDa. TAD = C-terminal transactivation domain. Pull downs have been replicated at least twice

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