Figure 6.
Hjv increases γ-secretase-like protease-cleaved Neo1 in the membrane. (A) Representative images of western blot analysis for Neo1, Tfr2, fHjv, and fHjvA183R in the liver membrane preparation (250 µg protein) of WT mice, PBS-injected Hjv−/− mice (–), and Hjv−/− mice transduced with AAV8-fHjv or fHjvA183R. qRT-PCR analysis of hepatic Neo1 (B), Hjv (C), and hepcidin (D) mRNA from mice in panel A. Data shown are means ± SD. (E) Serum iron (Fe) assay for mice in panel A. Each group consists of at least 5 animals. The means ± SD are presented. One-way ANOVA and Tukey’s posttests were used to analyze the data. ***P < .001. (F) PNGase F digestion. Liver membrane extracts (∼250 µg protein) from Hjv−/− mice transduced with AAV8-fHjv or fHjvA183R were subjected to PNGase-F digestion, followed by immunodetection with a horseradish peroxidase–conjugated anti-FLAG antibody. An image with 2 animals for each group was presented.

Hjv increases γ-secretase-like protease-cleaved Neo1 in the membrane. (A) Representative images of western blot analysis for Neo1, Tfr2, fHjv, and fHjvA183R in the liver membrane preparation (250 µg protein) of WT mice, PBS-injected Hjv−/− mice (–), and Hjv−/− mice transduced with AAV8-fHjv or fHjvA183R. qRT-PCR analysis of hepatic Neo1 (B), Hjv (C), and hepcidin (D) mRNA from mice in panel A. Data shown are means ± SD. (E) Serum iron (Fe) assay for mice in panel A. Each group consists of at least 5 animals. The means ± SD are presented. One-way ANOVA and Tukey’s posttests were used to analyze the data. ***P < .001. (F) PNGase F digestion. Liver membrane extracts (∼250 µg protein) from Hjv−/− mice transduced with AAV8-fHjv or fHjvA183R were subjected to PNGase-F digestion, followed by immunodetection with a horseradish peroxidase–conjugated anti-FLAG antibody. An image with 2 animals for each group was presented.

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