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. 2018 Jul 20;28(9):918–933. doi: 10.1038/s41422-018-0070-2

Fig. 7.

Fig. 7

Circulating apoptotic bodies rescued impaired MSCs in MRL/lpr mice. a Scheme illustrating parabiosis. MRL/lpr-MRL/lpr, MRL/lpr-GFP, and MRL/lpr-WT parabiosis models were used. b Compared to the control MRL/lpr-MRL/lpr parabiosis model, the amount of apoptotic bodies was significantly increased in MRL/lpr mice in the MRL/lpr-GFP parabiosis model. Flow cytometric analysis confirmed that the number of apoptotic bodies was increased in the bone marrow of MRL/lpr mice in the MRL/lpr-GFP parabiosis model. c Flow cytometric analysis showed that GFP and Annexin V double-positive apoptotic bodies were detected in the bone marrow of MRL/lpr mice in the MRL/lpr-GFP parabiosis model. The total number of Annexin V-positive apoptotic bodies was increased in MRL/lpr mice in the MRL/lpr-MRL/lpr parabiosis model. d Immunofluorescent staining showed that CD105-positive cells engulfed GFP apoptotic bodies in the bone marrow of MRL/lpr mice in the MRL/lpr-GFP parabiosis model. e Western blot showed that the levels of RNF146 and active-β-catenin were increased and the levels of Axin1 were decreased in MRL/lpr MSCs in the MRL/lpr-WT parabiosis model at 4 weeks post-parabiotic surgery. f BrdU labeling and continuous passage assay showed that MRL/lpr MSCs from the MRL/lpr-WT parabiosis model had increased proliferation and population doubling rates compared to MRL/lpr MSCs from the MRL/lpr-MRL/lpr parabiosis model. g MRL/lpr MSCs from the MRL/lpr-WT parabiosis model showed increased capacities to form mineralized nodules under the osteogenic inductive conditions, assessed by alizarin red staining (n = 5), and increased expression levels of ALP and Runx2, assessed by Western blot, when compared to MRL/lpr MSCs from the MRL/lpr-MRL/lpr parabiosis model. h MRL/lpr MSCs from the MRL/lpr-WT parabiosis model showed increased capacities to form new bone when implanted into immunocompromised mice compared to MRL/lpr MSCs from the MRL/lpr-MRL/lpr parabiosis model. i MRL/lpr MSCs from the MRL/lpr-WT parabiosis model showed increased capacities to differentiate into adipocytes under the adipogenic inductive conditions, as assessed by Oil red O staining (n = 5), and increased expression levels of PPAR-γ and LPL, as assessed by Western blot, when compared to MRL/lpr MSCs from the MRL/lpr-MRL/lpr parabiosis model. All results are representative of data generated in three independent experiments. Error bars represent the S.D. from the mean values. ***P < 0.001; **P < 0.01; *P < 0.05. Scale bar, 10 μm (d), 50 μm (h)