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. 2017 Nov 30;10(1):63–75. doi: 10.15252/emmm.201708168

Figure 4. The p.R119C and p.R38C THPO proteins are stable as the wild‐type THPO.

Figure 4

  1. Representative images of immunoblotting of HEK293T cells transfected with wild‐type or mutant (p.R119C and p.R38C) THPO‐expressing vectors. Cell lysates were prepared just before treatment with cycloheximide (CHX) (time 0) and 8, 24, and 48 h after the addition of CHX in order to block de novo protein synthesis (CHX chase assay). DMSO alone was used as control in the WT sample. GAPDH was used as loading control.
  2. Densitometric analysis of the bands obtained in n = 3 independent experiments. THPO amount was measured as THPO/GAPDH ratio and expressed as the percentage of the amount measured at time 0 in each condition (Relative protein, %). Of note, the expression of wild‐type THPO after CHX treatment was significantly lower compared with DMSO alone at each time point (P < 0.05, two‐tailed Student's t‐test), confirming that protein synthesis was efficiently blocked by CHX treatment. Data are presented as mean ± SD. The exact P‐values are reported in Appendix Table S3.

Source data are available online for this figure.