TABLE 1.
Plasmids used in this study
aa is amino acid. HA signal = MAIIYLILLFTAVRG Ig signal = MNFGFSLIFLVLVLKG CamLinker = PQPQPKPQPKPEPEGS PreSci = LEVLFQGP TEV = ENLYFQ.
| Plasmid | Designation | Protein encoded | Vector |
|---|---|---|---|
| ps193 | FLAG-hTNF | HA signal-FLAG-GPGQVQLQ-hTNF (aa 85–263) | PCR3 |
| ps420 | hTNFR1-Fc | hTNFR1 (aa 1–217)-VD-hIgG1 (aa 245–470) | PCR3 |
| ps515 | EGFP | Enhanced green fluorescent protein | PCR3 |
| ps645 | FLAG-hLTa | HA signal-FLAG-GPGQVQLQVD-hTNF (aa 43–205) | PCR3 |
| ps739 | hBCMA-Fc | Ig signal-EVKLVPRGS-hBCMA (aa 2–54)-VD-hIgG1 (aa 245–470) | PCR3 |
| ps897 | hTACI-GPI | HA signal-LD-hTACI (aa 2–160)-VD-hTRAILR3 (aa 157–269) | PCR3 |
| ps1196 | Fc-hBAFF | HA signal-LD-hIgG1 (aa 245–470)-RS-CamLinker-PreSci-GSLQ-hBAFF (aa 137–285) | PCR3 |
| ps1377 | pMSCV-puro | Modified pMSCV-puro (<zco;clontech>Clontech) with HindIII-BglII-EcoRI-NotI-XhoI-HpaI-ApaI cloning sites | ps1377 |
| ps1406 | Fc-hLTb | HA signal-LD-hIgG1 (aa 245–470)-RS-CamLinker-PreSci-GSLQ-hLTb (aa 66–244) | PCR3 |
| ps1467 | hBCMA-GPI | Ig signal-VQCEVKLVPRGS-hBCMA (aa 2–54)-VD-hTRAILR3 (aa 157–269) | PCR3 |
| ps1589 | hBAFFR-GPI | HA signal-LE-hBAFFR (aa 2–71)-EFGSVD-hTRAILR3 (aa 157–269) | PCR3 |
| ps1799 | mLTbR-Fc | mLTbR (aa 1–220)-VD-PreSci-hIgG1 (aa 245–470) | PCR3 |
| ps2297 | mBAFFR-Fc | HA signal-LD-mBAFFR (aa 2–70)-VD-hIgG1 (aa 245–470) | PCR3 |
| ps2308 | hBAFFR:Fas | HA signal-LE-hBAFFR (aa 2–71)-EFGSVD-hFas (aa 169–355) | ps1377 |
| ps2309 | hBCMA:Fas | Ig signal-VQCEVKLVPRGS-hBCMA (aa 2–54)-VD-hFas (aa 169–335) | ps1377 |
| ps2455 | hTACI:Fas | HA signal-L-hTACI (aa 1–118)-VD-hFas (aa 169–335) | ps1377 |
| ps2642 | FLAG-hLT-ααα | HA signal-FLAG-GPGQVQLQVD-hLTa (aa 58–205)-GGGGS-hLTa (aa 58–205)-GGGGS-hLTa (aa 58–205) | PCR3 |
| ps2643 | FLAG-hLT-ααβ | HA signal-FLAG-GPGQVQLQVD-hLTa (aa 58–205)-GGGGS-hLTa (aa 58–205)-GGGGS-hLTb (aa 83–244) | PCR3 |
| ps2644 | FLAG-hLT-ββα | HA signal-FLAG-GPGQVQLQ-hLTb (aa 83–244)-GGGGS-hLTa hLTb (aa 83–244)-GGGGS-hLTa (aa 58–205) | PCR3 |
| ps2645 | FLAG-hLT-βββ | HA signal-FLAG-GPGQVQLQ-hLTb (aa 83–244)-GGGGS-hLTa hLTb (aa 83–244)-GGGGS-hLTb (aa 83–244) | PCR3 |
| ps2680 | Fc-BBB | HA signal-LD-hIgG1 (aa 245–470)-RS-CamLinker-LQ-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285) | PCR3 |
| ps2826 | Fc-hAPRIL | HA signal-LD-hIgG1 (aa 245–470)-RS-CamLinker-GSLQ-hAPRIL (aa 115–250) | PCR3 |
| ps2876 | Fc-AAA | HA signal-LD-hIgG1 (aa 245–470)-RS-CamLinker-LQ-hAPRIL (aa 112–250)-GGGGS-hAPRIL (aa 112–250)-GGGGS-hAPRIL (aa 112–250) IRES GFP | PCR3 |
| ps2877 | Fc-BAA | HA signal-LD-hIgG1 (aa 245–470)-RS-CamLinker-LQ-hBAFF (aa 140–285)-GGGGS-hAPRIL (aa 112–250)-GGGGS-hAPRIL (aa 112–250) IRES GFP | PCR3 |
| ps2879 | FLAG-AAA | HA signal-FLAG-GPGQVQLQ-hAPRIL (aa 112–250)-GGGGS-hAPRIL (aa 112–250)-GGGGS-hAPRIL (aa 112–250) IRES GFP | PCR3 |
| ps2883 | Fc-ABB | HA signal-LD-hIgG1 (aa 245–470)-RS-CamLinker-LQ-hAPRIL (aa 112–250)-GGGGS-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285) IRES GFP | PCR3 |
| ps2886 | FLAG-ABB | HA signal-FLAG-GPGQVQLQ-hAPRIL (aa 112–250)-GGGGS-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285) IRES GFP | PCR3 |
| ps2889 | FLAG-BAA | HA signal-FLAG-GPGQVQLQ-hBAFF (aa 140–285)-GGGGS-hAPRIL (aa 112–250)-GGGGS-hAPRIL (aa 112–250) IRES GFP | PCR3 |
| ps2890 | FLAG-BBB | HA signal-FLAG-GPGQVQLQ-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285) IRES GFP | PCR3 |
| ps3413 | hTACI-Fc | Modified Ig signal (METDTLLLWVLLLWVPGVHG)-hTACI (aa 31–110) hIgG1 (aa 245–470; L258E, A353S, P354S) | PCR3 |
| pcDNA3.1_AAB | AAB | Ig signal-His6-FLAG-TEV-GS-hAPRIL (aa 111–250; T126A)-GGGGS-hAPRIL (aa 111–250; T126A)-GGGGS-hBAFF (aa 140–285) | pcDNA3.1 |
| pcDNA3.1_ABA | ABA | Ig signal-His6-FLAG-TEV-GS-hAPRIL (aa 111–250; T126A)-GGGGS-hBAFF (aa 140–285)-GGGGS-hAPRIL (aa 111–250; T126A) | pcDNA3.1 |
| pcDNA3.1_BAA | BAA | Ig signal-His6-FLAG-TEV-GS-hBAFF (aa 140–285)-GGGGS-hAPRIL (aa 111–250; T126A)-GGGGS-hAPRIL (aa 111–250; T126A) | pcDNA3.1 |
| pcDNA3.1_BBA | BBA | Ig signal-His6-FLAG-TEV-GS-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285)-GGGGS-hAPRIL (aa 111–250; T126A) | pcDNA3.1 |
| pcDNA3.1_BAB | BAB | Ig signal-His6-FLAG-TEV-GS-hBAFF (aa 140–285)-GGGGS-hAPRIL (aa 111–250; T126A)-GGGGS-hBAFF (aa 140–285) | pcDNA3.1 |
| pcDNA3.1_ABB | ABB | Ig signal-His6-FLAG-TEV-GS-hAPRIL (aa 111–250; T126A)-GGGGS-hBAFF (aa 140–285)-GGGGS-hBAFF (aa 140–285) | pcDNA3.1 |