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Fig. 3. Mapping of the JIL-1 interaction domain with lamin Dm₀. (A) The truncated COOH-terminal JIL-1 GST-fusion protein constructs used for domain mapping. The JIL-1 COOH-terminal domain can be divided into predominantly acidic and basic regions, with the basic region containing a predicted globular domain. (B) S2 cell lysate was incubated with the various truncated JIL-1 GST-fusion protein constructs shown in A or with a beads-only control and pelleted with glutathione-agarose beads. Interacting protein(s) were fractionated by SDS-PAGE, western blotted, and probed with the lamin mAb HL1203. Unincubated S2 cell lysate was included as a control (lane 6). The CTD, CTD-B and CTD-G were able to pull down the 76 kDa lamin protein (lanes 1, 3 and 4) also detected in the cell lysate (lane 6), whereas no interaction was observed with the CTD-A construct (lane 2) or with the beads-only control (lane 5). This defined the globular domain in the basic region as sufficient for mediating interactions with lamin Dm₀. (C) Immunoblot of the input GST-fusion proteins used for the pull-down experiments in (B) detected with the anti-GST mAb 8C7. The relative migration of molecular weight markers is indicated to the right in kDa.

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