International Journal of Stem Cells : eISSN 2005-5447

Fig. 4.

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Fig. 4. DEP-induced cFos expression inhibits the immunomodulatory function of WJ-MSCs. WJ-MSCs were transfected with control siRNA (si-cont) or cFos siRNA (si-cFos) for 4 hr and then were stimulated with 10 μg/ml DEP. After 12 hr, knockdown of cFos was confirmed by qRT-PCR (a) and western blot (b). (c∼e) After 48 hr of incubation, cells were mitotically inactivated and co-cultured with MNCs as previously described. (c) Representative histogram of total lymphocyte proliferation. (d) Proliferation of total, CD4+ T, and CD8+ T lymphocytes was quantified. (e) Number of cell divisions of total, CD4+ T, and CD8+ T lymphocytes was quantified based on dilution of CFSE. (f) si-cFos-transfected WJ-MSCs were stimulated with DEP for 24 hr and then cultured with osteogenic differentiation medium for 14 days. Calcium deposits were stained with Alizarin Red S. Light microscopy images of osteocytes show representative images from three independent experiments (Scale bar: 100 μm). Absorbance of calcium deposits was measured using a spectrophotometer. (g) Transcription of IL-1β, IL-6, IL-8, IL-18, and NLRP3 was determined by qRT-PCR and normalized to β-actin transcription. Data are presented as mean±S.D. of triplicate experiments (*p<0.05; **p<0.01; ***p<0.001).
International Journal of Stem Cells 2022;15:203-16
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