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. 2009 Oct 8;284(49):34433–34443. doi: 10.1074/jbc.M109.036202

FIGURE 1.

FIGURE 1.

Down-regulation of seladin-1 does not significantly affect APP processing in SH-SY5Y human neuroblastoma cells stably overexpressing APP751 (SH-SY5Y-APP751). A, quantitative PCR was used to assess seladin-1 knockdown efficiency at the mRNA level using two different seladin-1 siRNA concentrations (siSel, 0.65- and 1.3-μg total amounts of siRNA used in transfections) in SH-SY5Y-APP751 cells. On average, a 60% decrease in GAPDH-normalized seladin-1 mRNA levels was observed with both siRNA concentrations. B, APPim, APPm, and total APP and APP CTF (C83 and C99) levels were determined using Western blotting with APP C-terminal antibody (A8717). Blots were reprobed with antibodies against seladin-1, GGA3, and GAPDH. On average, a 40% decrease in GAPDH-normalized seladin-1 protein levels was detected. The GAPDH-normalized levels of different forms and fragments of APP or GGA3 were not significantly altered. ***, p < 0.001; **, p < 0.01, n ≥ 4, S.D. Ctrl, control.