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  Indian J Med Microbiol
 

Figure 4: Effect of chlorogenic acid on extracellular signal-regulated kinase ½ phosphorylation and cell cycle regulators in C6 glioma cells. C6 glioma cells were seeded in 100 mm dishes and incubated with serum-free medium for 24 h. The cells were treated with chlorogenic acid (100 and 300 μM) for 24 h. (a) Cell lysates were separated on 12% acrylamide gels and the level of ERK½ phosphorylation was measured by chemiluminescence. (b) The graphs are based on the band intensities from panel A. The intensity obtained from untreated cells was considered to be 100%. The data are presented as mean ± standard deviation (n = 3). (c-e) After total RNA extraction, real-time polymerase chain reaction analysis was performed. The values of relative gene expression were determined by calculating the value of the Δcycle threshold (ΔCt), normalizing the average Ct value to the control gene, glyceraldehyde 3-phosphate dehydrogenase, and then calculating the 2-ΔΔCt values. For each gene, the level of expression in untreated cells was considered as 100%. The data are presented as mean ± standard deviation (n = 3). *P < 0.05 compared with untreated cells. CA: Chlorogenic acid; FBS: Fetal bovine serum; ERK; extracellular signal-regulated kinase

Figure 4: Effect of chlorogenic acid on extracellular signal-regulated kinase ½ phosphorylation and cell cycle regulators in C6 glioma cells. C6 glioma cells were seeded in 100 mm dishes and incubated with serum-free medium for 24 h. The cells were treated with chlorogenic acid (100 and 300 μM) for 24 h. (a) Cell lysates were separated on 12% acrylamide gels and the level of ERK½ phosphorylation was measured by chemiluminescence. (b) The graphs are based on the band intensities from panel A. The intensity obtained from untreated cells was considered to be 100%. The data are presented as mean ± standard deviation (<i>n</i> = 3). (c-e) After total RNA extraction, real-time polymerase chain reaction analysis was performed. The values of relative gene expression were determined by calculating the value of the Δcycle threshold (ΔCt), normalizing the average Ct value to the control gene, glyceraldehyde 3-phosphate dehydrogenase, and then calculating the 2<sup>-ΔΔCt</sup> values. For each gene, the level of expression in untreated cells was considered as 100%. The data are presented as mean ± standard deviation (<i>n</i> = 3). *<i>P</i> < 0.05 compared with untreated cells. CA: Chlorogenic acid; FBS: Fetal bovine serum; ERK; extracellular signal-regulated kinase