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A
Schematic diagram of NrCAM's domain structure and sequential proteolytic processing. NrCAM is firstly cleaved by furin, then by ADAM10, and finally by the γ‐secretase. The red antibody indicates the binding region of the N‐terminal NrCAM antibody. ICD (intracellular domain), CTFf (C‐terminal fragment created by furin cleavage), CTFα (C‐terminal fragment created by ADAM10 cleavage).
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B
Detection of soluble, cleaved NrCAM (sNrCAM) and full‐length, mature NrCAM (mNrCAM) and soluble APPα (sAPPα) in neuronal supernatants and lysates (prepared from E16 neurons), treated with GI254023x (5 μM), or solvent for 48 h.
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C
Detection of sAPPα, sNrCAM, and mNrCAM in ADAM10 cKO neuronal supernatants and lysates at 7 days in vitro (DIV7). The neurons prepared from floxed ADAM10 (ADAM10fl/fl) mice were infected with a lentivirus encoding improved Cre recombinase (iCre) or a control GFP lentivirus at DIV2. Conditioned media were collected for 48 h.
Data information: In (B and C), densitometric quantifications of the Western blots are shown on the right (**
P < 0.01; ***
P < 0.001; ****
P < 0.0001, two‐sided Student's
t‐test,
n = 6–8). Given are mean ± the standard error of the mean. The mean levels of solvent‐treated cells were set to 1. Representative Western blots are shown.
Source data are available online for this figure.