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. 2013 Sep 3;288(41):29934–29942. doi: 10.1074/jbc.M113.492520

FIGURE 2.

FIGURE 2.

Brain-specific expression of mouse and human GAREM2. A, expression of GAREM2 mRNA in various mouse (left panel) and human (right panel) tissues. Shown is RT-PCR of GAREM1 (center row) and GAREM2 (upper rows) with β-actin as the loading control (bottom rows). B, protein expression of GAREM2 (top panel), GAREM1 (center panel), and β-actin (bottom panel) in various mouse tissues. The total cell lysate (20 μg) was separated by SDS-PAGE and analyzed by immunoblotting with each antibody. Immunoprecipitated (IP) FLAG-GAREM proteins as the positive control were prepared from lysates from COS-7 cells that were transfected with empty vector (Vector) or expression vectors for overexpressing the recombinant proteins of FLAG-GAREM1 (FL-GAREM1) and FLAG-GAREM2 (FL-GAREM2), with β-actin as the loading control (bottom panel). C, GAREM2 (top panel), EGFR (center panel), and β-actin (bottom panel) in neuronal cell lines or cultured cells derived from brain. Immunoprecipitated FLAG-GAREM proteins as the positive control were prepared from lysates from COS-7 cells that were transfected with empty vector or expression vectors for FLAG-GAREM1 (FL-GAREM1). Total cell lysates (20 μg of protein of cell lysate and 5 μg of protein of mouse brain lysate) were separated by SDS-PAGE and immunoblotted with each antibody with β-actin loading control (bottom panel). Neuro2A, mouse neuroblastoma cells; SH-SY5Y, human neuroblastoma cells; DAOY, human medulloblastoma cells; G16-9, human glioma cells.