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. 2016 Sep 30;113(41):E6055–E6063. doi: 10.1073/pnas.1605818113

Fig. 4.

Fig. 4.

Interaction between ERdj5 and SERCA2 depending on [Ca2+]. (A) Redox states of ERdj5 under treatment with thapsigargin (Tg) or tunicamycin (Tu). Transfected FLAG-tagged ERdj5 was modified with mPEG2000-mal in HEK293T cells treated with Tg or Tu for the indicated amount of time. Double or single asterisks denote modified ERdj5 bands. (B) Twenty-four hours after cotransfection of FLAG-tagged SERCA2b and Myc-tagged ERdj5/WT into HEK293 cells, the cells were treated with Tg or ionomycin for the indicated amount of time. The percentage binding of ERdj5 was normalized to that in untreated cells. (C) FLAG-SERCA2b was captured with FLAG-conjugated beads in lysates of cells transfected with FLAG-SERCA2b. Lysates of cells transfected with Myc-tagged ERdj5 were adjusted to contain the indicated concentration of CaCl2. The adjusted cell lysates were incubated with FLAG-SERCA2b-bound beads for the pull-down assay. ERdj5-Myc bound to SERCA2b was detected by immunoblotting with an anti-Myc antibody. Binding of ERdj5 with SERCA2b was quantified and shown below the panel.