BioMed Research International / 2014 / Article / Fig 3

Research Article

Lung Cancer Stem Cell Lose Their Stemness Default State after Exposure to Microgravity

Figure 3

(a) ALDH assay performed on H460 cells by flow cytometry. The H460 were cultured in sphere medium and exposed to RPM for 24 and 48 h. Baseline fluorescence was established by inhibiting ALDH activity (with DEAB) (left) and used to identify ALDH positive cells (without DEAB). (b) The histogram shows the quantitative results obtained in three independent experiments. Data indicated a significant reduction of ALDH activity in H460 RPM-treated cells at 24 h (Student’s -test, ) and at 48 h (Student’s -test, ). (c) Evaluation of the relative expression of Oct-4 and Nanog by quantitative Real-Time PCR at 6 and 24 h. The results show a significant decrease of Nanog and Oct-4 in RPM-exposed cells at 6 h (Student’s -test, ) and at 24 h (Student’s -test, ). The data represent mean ± SD of three independent experiments. (d) Histograms represent the Oct-4 and Nanog mRNA expression in RPM-treated spheroids and in the same sample further exposed to normal gravity condition. The values were compared to these obtained after 24 h of RPM exposure. Preliminary data did not show strong differences between RPM and on ground growth after RPM-exposition.

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