Extended Data Fig. 2: TAgD-TVacOVA-mediated proteasomal degradation of OVA in BMDCs.
From: Antitumour vaccination via the targeted proteolysis of antigens isolated from tumour lysates
a) HPLC-MS data to verify the encapsulation of AHPC. b) Flow cytometry analysis of the internalization of different formulations by DC2.4 cells through analysing the fluorescence of GFP. c) and d) Quantitative analysis of co-localization of GFP signals (green) with pVHL signals (red). The coefficients are close to 1 if they are highly colocalized. e) The degradation kinetics of GFP-Cre in DC2.4 cells after incubation with AHPC-GFP-Cre LNPs for 3 hours. The cells pre-treated with free AHPC molecules are employed for comparison. GFP-Cre fluorescence is monitored over 24 hours. f) SDS polyacrylamide gel electrophoresis of OVA and AHPC-OVA. g) Fluorescence spectra of the mixture of AHPC-OVA and pVHL after 1-hour incubation. The mixture of OVA, AHPC, and pVHL was used as comparison. pVHL OVA, and pVHL were pre-labelled with FITC and RITC, respectively. h) Western blotting analysis of OVA levels in the cytoplasm in BMDCs after incubating with different LNP formulations for 3 hours. The OVA levels are evaluated after further 24-hour incubation. LNP/protein = 10/1, w/w. Data are presented as mean ± s.d. from n independent experiments (n = 3). Statistical significance was analysed by t test for c, and two-way ANOVA with Tukey’s test for d.
