Fig. 5

AMFR promotes K11-related polyubiquitination of APP. A Ubiquitination alteration of APP incubated in vitro with AMFR recombinant protein and E1, E2, and Ub at 37 °C for 1 h. Data presented are representative of three separate experiments. B APP ubiquitination in HEK293T-hAPP cells transfected with empty vector or His-AMFR along with HA-Ub by Co-IP assays. The protein expression levels of AMFR and APP in the cell lysates were confirmed. Data presented are representative of three separate experiments. C APP ubiquitination in SH-SY5Y-hAPP cells transfected with empty vector or His-AMFR along with HA-Ub by Co-IP assays. The protein expression levels of AMFR and APP in the cell lysates were confirmed. Data presented are representative of three separate experiments. D Schematic presentation of Ub and its mutants. E APP ubiquitination in HEK293T-hAPP cells transfected with empty vector or His-AMFR along with HA-Ub or its mutants (K11, K29, K33, K48 and K63) by Co-IP assays. Data presented are representative of three separate experiments. F APP ubiquitination in SH-SY5Y-hAPP cells transfected with empty vector or His-AMFR along with HA-Ub or its mutants (K11, K29, K33, K48 and K63) by Co-IP assays. Data presented are representative of three separate experiments. G APP ubiquitination in HEK293T-hAPP cells transfected with empty vector or His-AMFR along with HA-Ub or its mutant (K11R) by Co-IP assays. Data presented are representative of three separate experiments. H APP ubiquitination in SH-SY5Y-hAPP cells transfected with empty vector or His-AMFR along with HA-Ub or its mutant (K11R) by Co-IP assays. Data presented are representative of three separate experiments