Figure_5.tif (2.68 MB)

Stability analysis of PBase protein, facilitated by flow cytometry.

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posted on 2014-02-24, 03:36 authored by Jin-Bon Hong, Fu-Ju Chou, Amy T. Ku, Hsiang-Hsuan Fan, Tung-Lung Lee, Yung-Hsin Huang, Tsung-Lin Yang, I-Chang Su, I-Shing Yu, Shu-Wha Lin, Chung-Liang Chien, Hong-Nerng Ho, You-Tzung Chen

Hela cells were transfected with pTriEx-mPB-2A-eGFP and pTriEx-NP-mPB-2A-eGFP plasmids. (A) APC fluorescence via anti-His tag antibody staining indicated the presence of mPB or NP-mPB. Because the eGFP moiety was cotranslated with PBase variants, the GFP fluorescence intensity was used as a reference for the mPB and NP-mPB translation levels of successfully transfected cells. (B and C) Y-axis represents accumulated cell counts. Although the NP-mPB group had a much smaller percentage of cells that were APC+ (4.65%) than the mPB group (18.2%), the percentages of GFP+ cells were similar (26.7% vs. 32.7%). (D) The APC fluorescence intensity in GFP+ populations represented the protein stability. The ratio of mean APC fluorescence (a.u.) of NP-mPB to mPB was 302 to 671.