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. Author manuscript; available in PMC: 2021 Mar 3.
Published in final edited form as: Cell Metab. 2020 Mar 3;31(3):564–579.e7. doi: 10.1016/j.cmet.2020.02.001

Figure 2. Mycoplasma infection confers host cells resistance to NAMPT inhibitors by preventing NAD depletion.

Figure 2.

(A) Mycoplasma prevent NAMPTi-induced NAD and ATP depletion. Total cellular NAD (NADH + NAD+) and ATP levels were measured in uninfected (no bacteria) or mycoplasma-infected CRC119 cells treated with 100 nM STF118804 (STF) (n=3, values are normalized to the corresponding time zero controls and expressed as mean ±SD, *p<0.05).

(B) Mycoplasma prevent NAMPTi-induced inhibition of glycolysis (ECAR) and oxidative phosphorylation (OCR). Uninfected clean or mycoplasma-infected CRC119 cells were treated with 100 nM STF118804 (STF) for 48 hours. The basal extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were measured using Seahorse instrument (n=7–8, values are expressed as mean ±SD, *p<0.05).

(C) Mycoplasma prevent NAMPTi-induced mitochondrial NAD depletion. Relative levels of mitochondrial total NAD were measured after 24 hours treatment with 100 nM STF118804 (STF) or DMSO control (n=3, values are normalized to the uninfected DMSO control and expressed as mean ±SD, *p<0.05).

(D) Mycoplasma prevent NAMPTi-induced loss of mitochondrial membrane potential. Mitochondrial membrane potential loss (% TMRE-negative cells) was measured by flow cytometry after 24 hours treatment with 100 nM STF118804 (STF) or DMSO control. (n=3, values are expressed as mean ±SD, *p<0.05).

See also Figure S2C.