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. 2018 Aug 20;19(9):e45889. doi: 10.15252/embr.201845889

Figure 4. TMEM41B is required for mobilization and utilization of endogenous fatty acids.

Figure 4

  • A
    H4 Cas9 TMEM41B KO and NT control cells were pulsed with the fatty acid analog Red C12, chased in complete medium or upon serum deprivation, and stained with MitoTracker. Cells were imaged live with an automated CV7000 confocal microscope. Scale bar: 20 μm.
  • B
    Fatty acid Red C12 intensity was measured within MitoTracker signal, and ratio between SD and CM conditions was calculated. Data are presented as mean ± SD (n = 3 technical replicates).
  • C–E
    H4 Cas9 TMEM41B KO and NT control cells were plated in XF96 plates and analyzed using a Seahorse Bioscience XF96. (C) Basal oxygen consumption rates (OCR) and extracellular acidification rates (ECAR) were measured in cells grown in complete medium. OCR/ECAR ratio is also reported. Data are presented as mean ± SEM (n = 30–32 technical replicates) from one representative experiment. (D) OCR was measured in cells grown in substrate‐limited medium and treated in the absence or presence of Etomoxir (Eto). Oligomycin (OA), FCCP (FC), and a mixture of rotenone and antimycin A (RA) were added as indicated. Data are shown as mean ± SEM (n = 69 technical replicates). (E) Endogenous FA utilization was calculated by subtracting basal normalized OCR values of Eto‐treated cells from untreated cells. Data are shown as mean ± SEM (n = 69 technical replicates).