A plasmid encoding GFP was co-transfected as an internal transfection control. The experiments were conducted twice with similar results. The white, orange, and blue arrowheads mark PIGS-BE, Split-BE-N, and Split-BE-C, respectively. Rubisco staining indicates equal protein loading. Source data ...
The human WT, A53T or A30P mutant αsynS11 plasmids were injected into 1 cell stage WT eggs together with a plasmid expressing OMM-GFP1–10, IMS-GFP1–10 or mt-GFP1–10. For injections, all plasmids were diluted in Danieau solution (58 mM NaCl, 0.7 mM KCl, 0.4 mM MgSO...
Switch fusions were transiently expressed using the FreeStyle HEK293 expression system. Briefly, transfection was initiated when the cell density of FreeStyle 293 suspension cells reached 3 × 106/mL. Subsequently, 100 μg of the expression vector plasmid was diluted in Opti-MEM (Gibco, 31...
APRIL or 9E10-IgG4m (pre-incubated with Myc-APRIL) CAR-T cells were co-cultured with RPMI8226-GFP cells (b), while BAFF or 9E10-IgG4m (pre-incubated with Myc-BAFF) CAR-T cells were co-cultured with IM9-GFP cells (d). Fluorescent labels included Hoechst (blue), anti-PKC-θ (...
(residues 110 to 177) of the DiB mutants23as well as leucine zipper peptides with adjacent upstream or downstream portions of the vector from the pMRBad-Z-CspGFP and pET11a-Z-NspGFP plasmids, correspondingly, and the upstream portion of the pET11a-Z-NspGFP plasmid including His-tag. ...
with rapamycin-induced bands corresponding to the molecular weight of appropriately spliced protein detected via western blot analysis in all cases (Fig.2b, Supplementary Table1). It should be noted that a basal splicing signal appeared in the sample of NIN/CIC-GFP combination without rapamycin trea...
The plasmid pAG153 was generated by inserting the gene for FKBP (amplified using primers ag183 and ag184) into pAG152 using restriction enzymes BglII and BspEI. The plasmid pAG241 was constructed by Gibson assembly of two fragments obtained by amplification of the plasmid pAG153 with the ...
All ORFs were BP-recombined into pDONR plasmid and the yielded Entry vectors were sequence verified. Entry vectors were then LR-recombined into the split TEV destination vectors pTag4C_X-V2R-NTEV-tevS-GV or pcDNA3.1_X-CTEV, plasmids were described in detail before22,23, to obtain the ...
(b) In vivo solubility screen of 18 Pyrobaculum test proteins15 expressed in a ''sandwich'' configuration with N-terminal GFP10 and C-terminal GFP11 (GFP10-A-GFP11, A 5 protein of interest) from pTET-GFP10/11 plasmid assayed with either GFP1–10 (top) or GFP1–9 (bottom) expressed...
Cells were seeded at a concentration of 1 Â 105 cells per well of a six-well plate on the day before transfection, which was done using Lipofectamine 3000 (LifeTechnologies) following the manufacturer's protocol (cotransfection of BirA* construct and Flp coding plasmid pPGKFLPobpA (addgene ...