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Fig. 1. Lamin Dm₀ interacts with JIL-1 in pull-down assays. (A) S2 cell lysate incubated with JIL-1-NTD (aa 1-211) or JIL-1-CTD (aa 927-1207) GST-fusion protein constructs or with a beads-only control was pelleted with glutathione-agarose beads and the interacting protein(s) fractionated by SDS-PAGE, western blotted, and probed with the lamin mAb HL1203. Unincubated S2 cell lysate was included as a control (lane 1). Only the JIL-1-CTD construct was able to pull down the 76 kDa lamin protein (lane 4) also detected in the cell lysate, whereas no interaction was observed with the GST-only control (lane 2) or the JIL-1-NTD construct (lane 3). (B) Immunoblot of the input GST-fusion proteins (JIL-1-NTD and JIL-1-CTD) used for the pull-down experiments in A detected with the anti-GST mAb 8C7. (C) S2 cell lysate incubated with a lamin-GST fusion construct (aa 260-622) or a GST-only control or with beads-only was pelleted with glutathione-agarose beads and the interacting protein(s) fractionated by SDS-PAGE, western blotted, and probed with affinity purified JIL-1 antiserum. Unincubated S2 cell lysate was included as a control (lane 1). The lamin-GST fusion protein construct was able to pull down the 160 kDa JIL-1 protein (lane 4) also detected in the cell lysate while no interaction was observed with the GST-only control (lane 3) or with the beads-only (lane 2). (D) Immunoblot of the input GST-fusion protein (lamin-GST) and the GST control used for the pull-down experiments in C 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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