Figure 5.
Exogenous SAMHD1 expression in HuT78 cells reduces the levels of cFLIPS protein and mRNA. HuT78 vector control and SAMHD1-expressing cells were seeded at density of 2 × 104 cells per 6-cm dish containing 5 ml growth media on day 0 following trypan-blue exclusion method. (A) On day 7 post-seeding, cell lysates were collected and immunoblotting analysis was performed for cFLIP, HA (SAMHD1) and GAPDH (loading control) (left panel). Densitometry was performed using ImageJ to quantify the relative cFLIPL and cFLIPS protein levels (right panel). ***, p < 0.001 (B) On day 7 post-seeding, total RNA was isolated from HuT78 vector or SAMHD1-expressing cells and qPCR analysis was performed to quantify the relative cFLIPL and cFLIPS mRNA levels. GAPDH mRNA levels were quantified as internal control. All the data presented are representative of 3 independent experiments. *, p < 0.05.