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Fig. 4 | Molecular Neurodegeneration

Fig. 4

From: The Alzheimer’s disease-linked protease BACE1 modulates neuronal IL-6 signaling through shedding of the receptor gp130

Fig. 4

BACE1 cleaves gp130 in neurons. a Immunoblots of conditioned media (CM) and lysates (LYS) of primary neurons transduced with two different gp130 shRNAs or a scrambled (SCR) shRNA as control. Sgp130 was pulled-down from the conditioned medium. Full-length gp130 and SEZ6 in the lysates as well as secreted soluble gp130 (sgp130) and soluble SEZ6 (sSEZ6) were detected. Actin and sNrCAM serve as loading controls. The monomer and dimer bands of sgp130 are indicated. b ELISA to detect sgp130 in concentrated conditioned neuronal medium. Shown are mean ± SD from N = 3 biological replicates. One-way ANOVA with Dunnett’s multiple comparisons test. c Immunoblots against indicated proteins in conditioned media and lysates of neurons and astrocytes in the absence (DMSO) or presence (C3) of the BACE1 inhibitor C3. Actin and sNrCAM serve as loading controls. d-e Quantification of sgp130 monomer and dimer bands in the neuronal medium (N = 14 biological replicates), as well as full-length gp130 in neuronal (N = 17 biological replicates) and astrocyte lysates (N = 11 biological replicates) from immunoblots in c. Shown are mean ± SD. Unpaired Student t-test. f ELISA measurement of sgp130 in the concentrated conditioned medium from experiments in c. Shown are mean ± SD from N = 6 biological replicates. Unpaired Student t-test. g Immunoblots of conditioned media (CM) and lysates (LYS) of primary, floxed BACE1 neurons transduced with a control lentivirus (Ctrl) or a Cre-expressing lentivirus (BACE1−/−). Sgp130 was pulled-down from the conditioned medium. Full-length gp130 and SEZ6 in the lysates as well as secreted soluble gp130 (sgp130) and soluble SEZ6 (sSEZ6) were detected. Actin and sNrCAM serve as loading controls. The monomer and dimer bands of sgp130 are indicated. h ELISA measurement of sgp130 in the concentrated conditioned medium from experiments in g. Shown are mean ± SD from N = 3 biological replicates. Unpaired Student t-test. ***: p < 0.001; ****: p < 0.0001

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