1. Fig. 2 Fas controls IL-1RA–sEV secretion in murine MSCs.
Fas controls IL-1RA–sEV secretion in murine MSCs. (A) Western blotting and semi-quantification of Fas expression in GMSCs and SMSCs (n = 3). (B) Secreted sEV-associated protein quantification from Fas-deficient MRL/lpr and wild-type (WT) control GMSCs (n = 5). (C) Western blotting and semi-quantification analysis of CD63, CD9, CD81, and IL-1RA from sEV from Fas-deficient MRL/lpr and WT control GMSCs. sEV-associated proteins from culture supernatants of equal numbers of cells in control and MRL/lpr GMSC groups were loaded (n = 3). (D) ELISA analysis of secreted IL-1RA from the culture supernatant from WT control and Fas-deficient MRL/lpr GMSCs (n = 3). (E) Western blotting and semi-quantification analysis of cytoplasmic IL-1RA from WT control and Fas-deficient MRL/lpr GMSCs (n = 3). (F) Immunocytofluorescent double staining of IL-1RA (green) and Fas (red) in WT control and Fas-deficient MRL/lpr GMSCs. Dashed lines indicate the cell edge. Scale bar, 20 μm. (G) ELISA analysis of IL-1RA secretion in the culture supernatant of MRL/lpr and Fas-overexpressing MRL/lpr GMSCs (n = 5). (H) Western blotting and semi-quantification analysis of cytoplasmic IL-1RA from MRL/lpr and Fas-overexpressing MRL/lpr GMSCs (n = 3). (I) ELISA analysis of IL-1RA secretion in the culture supernatant from WT control GMSCs treated with and without Fas small interfering (siRNA) (n = 5). (J) Western blotting and semi-quantification of cytoplasmic IL-1RA and Fas from WT control GMSCs treated with or without Fas siRNA (n = 3). All results are representative of data generated from at least three independent experiments. **P < 0.01, ***P < Error bars are means ± SD. All data were analyzed using independent unpaired two-tailed Student’s t tests.
Xiaoxing Kou et al., Sci Transl Med 2018;10:eaai8524
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