Figure 2
IFNγ induces the repression of MCJ in macrophages.
(A) RAW cells were stimulated with live B. burgdorferi (m.o.i = 25) or 100 ng/mL of LPS for 16 h and analyzed by immunoblotting for MCJ protein levels. Actin levels were determined to ensure equal loads. (B) BMMs were stimulated with live B. burgdorferi for 16 h and analyzed for MCJ mRNA levels by qRT-PCR. The data shown correspond to the mean ± SE of 3 points per group. (C) RAW cells (RAW), BMMs, Hepa 2–7 cells (Hepa) or CD8+ T cells (CD8) were stimulated with 100 ng/mL of IFNγ for 24–48 h, followed by the analysis of MCJ protein levels by immunoblotting. GAPDH levels were determined to ensure equal protein loads. (D) RAW cells were stimulated for 72 h with 100 ng/mL of IFNγ in 8-well chamber slides, washed and stained for intracellular MCJ. The slides were analyzed by ApoTome fluorescence microscopy. (E) RAW cells stimulated with IFNγ were analyzed for the levels of the mitochondrial protein, VDAC1, by immunoblotting. (F) RAW, BMMs and Hepa cells were stimulated with 100 ng/mL of IL-6 for 24 h, followed by their analysis for MCJ protein content by immunoblotting.
