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. 2020 Nov 12;15(11):e0241685. doi: 10.1371/journal.pone.0241685

Fig 3. CBS dependent regulation of H2S and expression of pluripotency gene, OCT4 in 3D spheroids.

Fig 3

A. H2S released by HDF after 6 day of culture. B. H2S stained with 5 μM H2S fluorescent probe, HSN2. C. qPCR of expression of H2S-synthesizing enzymes, CBS, CSE, MPST. β-ACTIN was used as a housekeeping control. Cells were transfected with CBS siRNA or scrambled (Scr) control siRNA. D. qPCR of expression of CBS. E. Amount of H2S in culture media. F. Amount of lactic acid in culture media. G. qPCR of expression of OCT4 and NANOG after 72 h of culture. β-ACTIN was used as a housekeeping control. Data are presented as the mean ± SD from 3–6 independent experiments. Statistical significance is shown as * (p≤0.05), ** (p ≤0.005), and *** (p ≤0.0005) in 3D as compared to 2D cultured cells.