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. 2020 Mar 16;11(3):188. doi: 10.1038/s41419-020-2378-3

Fig. 1. Lentiviral transduction of BM-MSCs with shPHD2-GFP specifically reduced PHD2 expression.

Fig. 1

a The experimental design is depicted for in vivo and in vitro experiments. For the in vivo experiments, newborn Sprague-Dawley rats delivered by caesarean section were not exposed (control) or exposed to experimental stresses (NEC), followed by a course of i.p. injection with control medium (DMEM-F12) or conditioned medium from GFP-transduced BM-MSCs (MSC-CM) or shPHD2-GFP-transduced BM-MSCs (PHDMSC-CM). Pups were either killed on days 1, 4, 7 for structural and functional examination or were monitored for survival status throughout the 7 day course of the experiment. For in vitro experiments, control rat intestinal epithelial IEC-6 cells or cells exposed to experimental stresses were cultured for 7 days. In parallel, IEC-6 cells were co-cultured in six-well plates with unconcentrated MSC-CM or PHDMSC-CM, and the proliferation and apoptosis were evaluated every other day as shown. b BM-MSCs with lentiviral transduction of BM-MSCs with GFP or shPHD2-GFP constructs. Scale bar = 100 μm. c Representative blots of PHD2, PHD1 and PHD3 expression in BM-MSCs with or without shPHD2. d Flow cytometric analysis shows that PHDMSC were positive for CD29, CD44 and negative for CD34, CD45, suggesting that the gene transfer procedure did not alter the MSCs phenotype.