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. 2018 May 29;46(13):6627–6641. doi: 10.1093/nar/gky451

Figure 1.

Figure 1.

SisPINA has physical interaction with SisHjm. Analysis of the interaction between SisPINA and SisHjm by pull-down and gel filtration. (A) Recombinant SisPINA or SisPINA (1–492) (without His-tag) was incubated with N-terminal His-tagged SisHjm. Inputs and elution fractions were analyzed by SDS-PAGE with Coomassie blue staining. M, molecular size markers. ‘–’, no protein, ‘+’, protein added. Molecular masses of standard proteins are indicated at the left of each panel. (B) Gel filtration profile of Hjm (red), SisPINA (black), Hjm/SisPINA mixture (blue), and protein marker ferritin (cyan, 440 kD) (GE Healthcare, UK). The protein sample (500 ul) of SisPINA (430 μg), Hjm (230 μg), or their mixture was loaded onto the Superdex 200 column which was equilibrated with the buffer containing 25 mM Tris–HCl, pH 8.0, 200 mM NaCl, and 5% glycerol. (C) SDS-PAGE analysis of the elute fractions in (B). The fractions (1 ml each) were collected and analyzed.