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Lysate PS1 IP NCT IP APH-1 IP PEN-2 IP N.R. Sera IP TMP21 17
(SHSY-5Y cells) 17 Flag- (HEK293 cells) (Mouse brain)
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TMP21 NCT PS1-NTF APH-1 PEN-2 Fraction No. 1 2 3 4 5 6 7 8 9 10 11 12
29 69 150 200 443 Marker (kDa): 17 34 105 Mr. (kDa)
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Mr. (kDa): TMP21 APH-1 NCT PEN-2 PS1-FL PS1-NTF Native cells PEN-2 Knockdown cells PS1 & PS2 double Knockout cells
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TMP21 NCT GM130 1 2 3 4 5 6 7 8 9 10 11 12 biotinylation
mature immature 1 2 3 4 5 6 7 8 9 10 11 12 biotinylation lysate IP unbound with avidin no biotinylation
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Ab40 Ab42 TMP21 Aβ assays in whole cell Aβ assays in cell
CHAPSO solubilized Membrane fraction Conditioned media Microsomal membranes Ab40 peptide control Ab42 peptide control Ab40 Ab42 TMP21 Aβ assays in whole cell Aβ assays in cell free/ g-secretase preps over-expression Empty vector Tmp21
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TMP21 Control siRNA TMP21 siRNA Ab
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Ab production (fmol/ml) mock p24a over-expression
100 200 Ab production (fmol/ml) mock p24a over-expression Ab40 Ab42 overexpression Empty vector p24a PS1-NTF NCT APH-1 TMP21 PEN-2 MW 34 105 16 17 7 Aβ assays in cell free/ g-secretase preps Aβ assays in whole cell
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TMP21-Flag Untransfected TMP21-Flag
PS1/PS Compound E Cell type HEK HEK HEK PS1/PS2 PS1/PS2 PS1/PS2 PS1/PS2 D-KO WT D-KO WT Mr (kDa) 23 17 7 Full-length TMP21
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APP Control siRNA TMP21 siRNA Mock Mat Imm Chase Time (min):
Control siRNA TMP21 siRNA Mock APP Mat Imm
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+ + + + + + - TMP21 BACE APP Nct Cell lysate Avidin precipitation
Mock Con TMP21 Mock Con TMP21 Mock siRNA Biotinylation 23 105 53 MW
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Negative Ab40 Ab42 Tmp21 Duplex A Duplex B Duplex C Duplex D
Tmp21pool Duplex A Duplex B Duplex C Duplex D Ab42 peptide Ab40 peptide
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