Figure 1 | Scientific Reports

Figure 1

From: The impact of hypoxia and oxidative stress on proteo-metabolomic alterations of 3D cholangiocarcinoma models

Figure 1

Formation of 3D MCSs using five CCA and cholangiocyte cell lines. (a) The process of agarose-coating (U-surface), showing the coating procedures. The illustration was created with BioRender.com. (b) Four CCA cell lines (KKU-055, KKU-100, KKU-213C, KKU-213A) and one cholangiocyte cell line (MMNK-1) are shown. Each cell line is in a 2D monolayer culture system. (c) 3D cultures of 5 cell lines in the coated U-bottom plate, incubated for 48 h. (d) Cells transferred back to the flat-bottom plate, showing only KKU-055 and KKU-100 scattered to single cells, while the rest maintain intact spherical shape as 3D MCS, scale bar represents 200 µm. (e and f) Western blot analysis of cell-cell adhesion molecules (E-cadherin, N-cadherin, and P-cadherin), mesenchymal marker (vimentin), enzymes involved in the glycolytic pathway (GLUT-1, GAPDH, and LDHA), enzymes related to TCA cycle (IDH1 and IDH2), and angiogenic factor (VEGF-C) express on 2D normoxia vs. 2D hypoxia (e) and 2D normoxia vs. 3D spheroid (f) of CCA and cholangiocyte cell lines. Quantitative analysis of protein (normalized to β-actin) is shown in supplement S1. Data are means ± S.D. of at least three independent experiments. *P < 0.05, **P < 0.01 and ****P < 0.01.

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