Desquamated Epithelial Cells of Unstimulated Human Whole Saliva Express Both EGF Transcript and Protein

Figure 6

EGF gene expression is lower in growth-stimulated than in growth-restricted HaCaT cells. (a–c) Cells were incubated in the FBS-free culture medium for 8 h to standardize the experimental conditions and then stimulated with either low (0.1%) or high (10%) concentration of FBS for 24 h representing growth-restricted and growth-stimulated conditions, respectively. EGF mRNA levels were determined by qPCR using the delta-delta Ct method (a). Cells were stained with trypan blue, and the number of cells was counted using an automatic cell counter (b). Total protein concentration in cell lysates was investigated using a Bio-Rad DC protein assay kit (c). Experiments were performed three times in duplicate or triplicate. Values are presented as means ± SEM for three experiments in each group. Statistical significance was calculated using Student’s two-tailed t test for single comparison between the two groups (0.1 vs. 10% FBS) as appropriate. represents , and represents .

(a)

(b)

(c)