Fig. 4.

Tissue distribution and intracellular localization of TMEM86a. A: Wild-type mice (N = 5/group) were pharmacologically dosed with GW3965 for 4.5 days (20 mg/kg, twice daily). Organs were collected and gene expression determined by qPCR. BMDMs were isolated independently. B: Predicted membrane topology of mouse TMEM86a. Conservation with TMEM86b and residues predicted to form the catalytic site are indicated. C, D: Total cell lysates from HEK293T cells expressing Myc-TMEM86a or TMEM86a-V5 were fractionated and immunoblotted as indicated. GM130 and GAPDH were used to distinguish the membrane and cytosolic fractions, respectively. E: BMDMs were transduced with lentivirus encoding Myc-TMEM86a and stained as indicated. DAPI was used to counterstain the nuclei and cells were imaged by confocal microscopy. The mean ± SEM are depicted. ∗P<0.05, ∗∗∗∗P < 0.0001.