Fig. 9: SCAP combined with IκBɑ-mediated Golgi apparatus translocation activates NF-κB. | Experimental & Molecular Medicine

Fig. 9: SCAP combined with IκBɑ-mediated Golgi apparatus translocation activates NF-κB.

From: Cholesterol-driven pathological astrocytic responses in diabetes-associated cognitive impairment through astrocytic SCAP accumulation and NF-κB–C3 signaling modulation

Fig. 9

a The astrocytes were analyzed for colocalization of BAP31 and Golgi 97 with IκBɑ in the basal state via confocal microscopic imaging. Scale bars, 10 μm. b, c The representative immunoblots (b) and the quantification (c) of astrocytic IκBɑ and p-IκBɑ after treatment with different concentrations of glucose (Gluc) (5.5 mM or 25 mM) or (100 μg/ml) LDL-C (n = 4 per group per experiment). d The astrocytes were treated with Gluc (25 mM) and LDL-C (100 μg/ml), and the colocalization of BAP31 and Golgi 97 with p-IκBɑ was analyzed. Scale bars, 10 μm. e The immunofluorescence staining of SCAPs and IκBɑ in astrocytes. Scale bars, 10 μm. f A schematic diagram of the mechanisms by which astrocytic SCAP activates IκBɑ/NF-κB. g The immunoblotting for immunoprecipitation with anti-SCAP, anti-IκBɑ and anti-NF-κB antibodies in astrocytes (n = 3). h, i The representative immunoblots (h) and the quantification (i) of astrocytic IκBa and p-IκBɑ in WTA and KOA treated with Gluc (25 mM) and LDL-C (100 μg/ml) (n = 4 per group per experiment). j The SCAP-KOA and control astrocytes (WTA) were treated with Gluc (25 mM) and LDL-C (100 μg/ml), and the colocalization of Golgi 97 with p-IκBɑ was analyzed in KOA and WTA. The data are expressed as the means ± s.e.m. The one-way ANOVA with Tukey’s multiple comparisons test was performed for c, and Student’s t-test (two-sided) was performed for i. n.s., nonsignificant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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