Journal: Molecular Metabolism

Article Title: Activated macrophages control human adipocyte mitochondrial bioenergetics via secreted factors

doi: 10.1016/j.molmet.2017.07.008

Figure Lengend Snippet: Effects of LPS/IFNγ- and IL10/TGFβ-activated primary MΦ-CM on bioenergetics of human SGBS and primary adipocytes . Primary human CD14 + monocytes from 4 healthy donors were cultured, differentiated to macrophages (naïve MΦs) and activated with LPS/IFNγ or IL10/TGFβ as described in . (A) Morphological changes detected by bright field microscopy (one representative experiment shown). (B) mRNA expression of CD163 and CD40 analyzed with ΔΔCT method using naïve MΦs as Calibrator (n = 4). (C) CD163 and CD40 mRNA expression presented as ratio. (B, C) *p < 0.05 vs. LPS/IFNγ-activated MΦ, ***p < 0.001 vs. LPS/IFNγ-activated MΦ, ###p < 0.001 vs. naïve MΦs. (D) mRNA expression of UQCRC2 and NDUFB8 analyzed with ΔΔCT method using naïve THP1 macrophages as Calibrator (n = 4). ( E–G) SGBS adipocytes were treated with either cell-free control media, LPS/IFNγ-activated or IL10/TGFβ-activated MΦ-CM collected from 3 different donors for 48 h as described in and analyzed as described in Figure 4 , Figure 5 . Data are normalized to cell number (DNA content) and are the mean + SEM of 5 independent experiments each performed in 2–8 wells condition. OCR and ECAR traces vs time are shown in ( Figure S4B and C ). (E) ATP-linked respiration of SGBS adipocytes after treatment for 48 h with indicated MΦ-CM. (F) ATP-linked respiration of SGBS adipocytes presented as relative change in OCR in percent of cell-free control. (G) ECARs due to glycolysis of SGBS adipocytes after treatment for 48 h with indicated MΦ-CM. (H – J) Human primary subcutaneous adipocytes were treated with either cell-free control media, LPS/IFNγ-activated or IL10/TGFβ-activated MΦ-CM from 3 different donors as described in . Bioenergetic profile was analyzed as described in Figure 4 , Figure 5 . Data are presented as fold change to the mean rates of cell-free control for each donor and are the mean + SEM of experiments from 3 adipocyte donors performed in 6–8 wells per condition and donor. OCR and ECAR traces vs time are shown in ( Figure S4E and F ) (H) ATP-linked respiration of human primary adipocytes after treatment for 48 h with indicated MΦ-CM. (I) ATP-linked respiration of human primary adipocytes presented as relative change in OCR in percent of cell-free control. (J) ECARs due to glycolysis of human primary adipocytes after treatment for 48 h with indicated MΦ-CM. (E)–(J) *p < 0.05 vs. LPS/IFNγ-activated MΦ-CM; **p < 0.01 vs. LPS/IFNγ-activated MΦ-CM; ***p < 0.001 vs. LPS/IFNγ-activated MΦ-CM; #p < 0.05 vs. cell-free control media.

Article Snippet: Human CD14 + blood monocytes from 4 healthy donors were purchased from Zenbio (Research Triangle Park, NC, USA) and differentiated with 10 ng/ml CSF1 for 7 days to adherent macrophages (MΦ) before same stimulation as for THP1 cells was performed.

Techniques: Cell Culture, Microscopy, Expressing, Control