Figure 1.

CoCl₂ affects the proliferation, migration, and colony formation of rat glioma cells in a hypoxic model. (a) The data of RT-PCR are presented. Histogram bars show the relative amount of mRNA transcribed from the hif1α gene in C6 cells 6 h after addition of CoCl₂; the data are normalized to the amount of actin mRNA. (b) Cell index data were obtained using the xCELLigence device. The graph shows the dynamics of the C6 cellular index placed in E-plates; CoCl₂ was added in concentrations of 50, 100, and 200 μM after cell attachment to the bottom; incubation time is indicated on the x-axis. Representative data from three independent experiments are shown. (c) Data on C6 cell motility data obtained using the xCELLigence system. The graph shows the dynamics of the cellular index measured in the upper wells of the Cell Invasion and Migration (CIM) plates of the xCELLigence system in a medium containing 50, 100, or 200 μM CoCl₂. The lower chambers of the CIM plates were filled with CoCl₂-free medium. The recording of cell migration through the microporous membrane of the CIM plates lasted 40 h. Representative data from three independent experiments are shown. (d) C6 cell colony formation efficacy in the presence of 50, 100, and 200 μM CoCl₂. (e) The result of colony number quantification from section (c) is presented. The histogram bars show the average normalized number of colonies based on three independent experiments. (f,g) Wound healing assay was performed with the aid of the JuLI Stage microscope and monitored by JuLI software. Cells were cultivated for 24 h in the presence of CoCl₂ in different concentrations before the monolayer was scratched. The wound healing was detected with microscopy (f) and quantified with the aid of JuLI software (g). (h) The results of the CytoTox96 assay. C6 cells were incubated with CoCl₂ for 24 h. Data presented as mean ± standard error of mean (SEM). Statistical significance is indicated as * p < 0.05 and ** p < 0.01.