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. 2020 Sep 29;18(10):498. doi: 10.3390/md18100498

Figure 9.

Figure 9

Expression of stemness and pluripotency biomarkers in R182 cell spheroids compared to 2D cultures. (A) RT-PCR gene expression analysis of stemness and pluripotency regulatory factors (Sox2, Nanog, Oct4, and KLF4) in 2D-cultured cells, 3D spheroids, and the isolated CD133+ CSCs from 3D spheroids. Bar graphs show the relative expressions of genes in cells grown in these cells. GAPDH was used as a housekeeping gene for RT-PCR data normalization. Results are expressed as mean ± SD (n = 3). (B) Flow cytometry comparative analysis of the expression of aldehyde dehydrogenase 1 family member A1 (ALDH1A1), a functional stem cell marker, in 2D and 3D cultures. Data represent the means ± SD of three independent experiments. *** p < 0.001 vs. 2D. # p < 0.05 and ## p < 0.01 vs. 3D spheroids.