Fig 1: Cell plasticity of MDA-MB-231 cells precultured in different cell culture configurations. (a) WB corresponding to the intracellular STC1 expression of “primed” MDA-MB-231 cells further cultured for 2 days on TCP under the same cell densities. a-Tubulin was used as a loading control. (b) Bar graph corresponding to secreted STC1 normalized to cell number. STC1 is measured by ELISA from the supernatant of “primed” MDA-MB-231 cells after 2 days of culture on TCP under the same cell densities and normalized to the cell number shown in the (e) bar graph. (c) Bar graph corresponding to STC2 secretion normalized to cell number. STC2 is measured by ELISA from the supernatant of “primed” MDA-MB-231 cells after 2 days of culture on TCP under the same cell densities and normalized to the cell number shown in the (f) bar graph. (d) Bar graph representing WB semiquantitative analysis of STC1 expression corresponding to STC1 expression after 2 days of cell culture and normalized to a-tubulin. (e) Average cell number determined by DNA quantification corresponding to each culture condition of STC1 experiments. (f) Average cell number determined by DNA quantification corresponding to each culture condition of STC2 experiments. Biological triplicates were used per experiment, and technical triplicates were used during the performance of ELISAs. Error bars represent SD. One-way ANOVA shows no statistical differences among all groups of “primed” cells once cultured for 2 additional days on 2D TCP. One-way ANOVA shows no statistical differences of cell number among all groups in the bar graph (e) corresponding to STC1 secretion experiments and the bar graph (f) corresponding to STC2 secretion experiments.
Fig 2: Expression of dormancy-related markers in MDA-MB-231 cells cultured in different configurations. (a) WB corresponding to the intracellular STC1 expression of MDA-MB-231 cells cultured for 8 days on either 2D TCP or 3D PA scaffolds. a-Tubulin was used as a loading control. (b) Bar graph corresponding to normalized to cell number STC1 secretion, measured by ELISA from the supernatant of cell cultures at t = 10 days. (c) Bar graph corresponding to normalized to cell number STC2 secretion measured by ELISA from the supernatant of cell cultures at t = 10 days. (d) Bar graph representing WB semiquantitative analysis of STC1 expression corresponding to normalized (to a-tubulin) STC1 expression after 8 days of cell culture on the various systems. (e) Average cell density corresponding to each culture condition of STC1 experiments. The cell number of each scaffold (data not shown) was used for the STC1 normalization corresponding to each scaffold. (f) Average cell density corresponding to each culture condition of STC2 experiments. The cell number of each scaffold was used for the STC2 normalization of each scaffold. Biological triplicates were used per experiment, and technical triplicates were used during the performance. Error bars represent SD, and * and ** denote P < 0.05 and P < 0.005, respectively.
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