Figure legends
Figure 1 . Schematic illustration of the generation of single
tumor spheroid.
Figure 2 . Physiological characterizations of the growth
dynamics of 3D MTSs over the culture age. (A) Optical imaging of 3D
MTSs. (B) Diameter of 3D MTSs. (C) Circularity of 3D MTSs. (D) Growth
curve of 3D MTSs. (E) The morphology and
microstructure. (F) Cell apoptosis
and cell cycle.
Figure 3 . The effect of 5-FU on 3D MTSs. (A) Cytotoxicity
assay. (B) Cell apoptosis and cell cycle. (C) Mitochondrial respiration
profile. (D) ROS measurement. The measured concentrations of
extracellular (E) glucose, (F) glutamine, (G) ammonium and (H) lactate.
Figure 4 . Transcriptional analysis of putative drug resistance
genes associated with HIF1A -induced signal pathways, glucose
metabolism, nucleotide metabolism, multidrug resistance-related geneLAPTM4B , ECM secretion,
tumor stem cell and signaling pathways related to cell growth and
proliferation.
Figure 5 . Proteomic
differences between cells in 2D monolayer and 3D MTS. (A)
Heatmap of differential proteins
between 2D monolayer and 3D MTS.
50 differential proteins in the
top KEGG pathways (sorted according to the -log10 p-value) were selected
to draw a heat map. (B) The top 20 KEGG pathways of proteome between 2D
monolayer and 3D MTS (sorted according to the -log10 p-value). (C)
Chord diagram of GO analysis.
The
top 6 GO terms with the number of differential proteins between
3~50 was selected by sorted according to the -log10
p-value in descending order. The relationship between the selected GO
term and the corresponding differential proteins were shown in the chord
diagram. (D) Protein-protein
interaction network of the top 25 proteins in connectivity degree. The
molecular chaperones related to protein folding, assembly, and secretion
exerted the most obvious effects, and the heat shock protein family was
dominant.
Figure 6 . Metabolomic differences between cells in 2D monolayer
and 3D MTS. (A) Heatmap of differential metabolites between 2D monolayer
and 3D MTS. 50 differential metabolites in the top KEGG pathways (sorted
according to the -log10 p-value) were selected to draw a heat map. (B)
The top 20 KEGG pathways of proteome between 2D monolayer and 3D MTS
(sorted according to the -log10 p-value).