Figure 3. Differential expression of the growth-related signal proteins and Rex-1 involvement in active growth of low oxygen/DHP-d exposed ATSC.

(A, B) The involvement of the JAK/STAT3 and MEK signal protein in active cell growth after de-differentiation of ATSC. For the confirmation of differentially expressed proteins following the de-ATSC, the cells lysates were subjected to SDS-PAGE analysis and transferred to nitrocellulose membranes. Optimally diluted antibodies were incubated with the membranes. The relative band intensities were determined using Quality-one 1-D Analysis software. (C) Prominent inhibition of de-ATSC growth by Rex1 siRNA and (D) Oct4 siRNA. Two complementary hairpin siRNA template oligonucleotides harboring the 21 nt target sequences of the human Rex1 were employed for transient transfection. Three separate Rex1 siRNAs and scrambled siRNAs with the same nucleotide content were assessed. For the detection of the inhibition of de-ATSC growth, we transfected Rex1 siRNA into de-ATSC counted dye-exclusive viable cells for 6 days. (E) Functional involvement of HIFs in de-ATSC proliferation, proliferation controlling signal protein, and stemness genes expression. (F) Schematic flow chart of the low oxygen/DHP-d induced ATSC proliferation signal pathway involving Rex1, Nanog, p53, p21, and c-myc gene expression and activation in the nucleus. Datas presented are presented as mean ±SD; n>5. * p % 0.05, and ** p % 0.01, Student's t test.