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. 2010 Apr 28;285(27):20664–20674. doi: 10.1074/jbc.M109.055608

FIGURE 3.

FIGURE 3.

Knockdown of TMEM59 and TMEM59L affects APP shedding. A, HEK293 cells were transfected with control siRNA or siRNAs against TMEM59 and TMEM59L. Endogenous APP and its cleavage products were detected as described in the legend to Fig. 2A. B, knockdown (KD) of TMEM59 and its homolog reduced the amount of secreted APP (APPsα and APPsβ) by about 30–40% compared with control cells. Quantification of blots shown in A. Given are mean ± S.E. of 4 independent experiments. *, p < 0.05, determined with t test. C, siRNA treatment reduced mRNA levels of TMEM59 and TMEM59L by about 70–80%, as determined by quantitative reverse transcription-PCR. Given are mean ± S.E. of 4 independent experiments. ***, p < 0.001, determined with t test. D, endogenous TMEM59 (antibody 93) was detected in permeabilized HEK293 cells treated with control siRNAs (con) but not in cells treated with siRNAs against TMEM59 and TMEM59L (KD). Scale bars = 10 μm. For a more detailed immunofluorescence analysis, see Fig. 5.