Copper Uptake in Mammary Epithelial Cells Activates Cyclins and Triggers Antioxidant Response

Figure 3

Real-time PCR revealed that Cu stimulated mild upregulation of cyclin D1 and cyclin B1 genes in MCF10A cells. Expression of cyclin genes in cells treated with 50 μM CuSO₄. The results were plotted and normalized with respect to respective untreated cells. The specificities of the primers for human glyceraldehyde 3-phosphate dehydrogenase (GAPDH), cyclin D1, and cyclin B1 were verified by real time PCR. Amplification plots, linear regression, and dissociation melting curves for (a–c) cyclin D1, (d–f) cyclin B1, and (g-h) GAPDH obtained with serial dilutions of cDNA (1, 1/3, 1/9, and 1/27). (j) Expression of cyclin D1 in Cu-treated MCF10A cells. Significant differences between untreated and Cu-treated cells are indicated by asterisks (, ).

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)