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C/EBP knockdown, for instance, decreased expression of all genes measured, including ATF4, which was also reflected in loss of ATF4 protein (Figure 2C-D)

Such RNA interfering agents include, but are not limited to, nucleic acid molecules (including RNA molecules homologous to a target biomarker gene or fragment thereof encompassed by the present invention), short interfering RNAs (sirnas), and small molecules that interfere with or inhibit expression of a target biomarker nucleic acid by RNA interference (RNAi)

For Immunofluorescence stainings, the tissue sections were fixed with 4% paraformaldehyde (PFA) for 10 min and permeabilized using 0.1% Triton X-100 for 10 min at room temperature, followed by blocking using 3% bovine serum albumin (BSA) for 1 h at room temperature

These processes are fundamental in biological systems, setting off a chain of reactions in organisms that utilize oxygen from the air for oxidation, producing energy in the form of ATP (Gulcin 2012