Supplementary materials

Figure S1. Characterization of PEG-PE micelle

(A) Transmission electron microscopy (TEM) image of PEG-PE micelles encapsulating OVA 250-264

peptide. Scale bar, 50 nm.

(B) Representative size distribution of PEG-PE micelle was measured by dynamic light scattering

(DLS) analysis.

Figure S2. Most of 200nm liposome stay in SCS region of LNs.

(A) Representative size distribution of liposome was measured by dynamic light scattering (DLS)

analysis.

(B) Transmission electron microscopy (TEM) image of liposome. Scale bar, 100 nm.

C57BL/6 Mice were s.c injected with RhoB (red) labeled 200nm liposome at tail base, and draining

inguinal LNs were isolated at 2 hours (C) or 6 hours (D) for visualization of its distribution.

Identification of stromal elements within LNs, with LYVE1(blue) signal demarcating the SCS. Box 1-

4 indicate enlarged areas of LN montage. Color of the word labels correspond to the colors of the

stains here.

Figure S3. PEG-PE micelle efficiently deliver OVA250-264 peptide into the cell cytosol.

DC2.4 cells were incubated with 10μM RhoB-labeled OVA250-264 or PEG-PE encapsulated RhoB-

OVA250-264(PEG-PE/OVA), respectively for 30 min. Subsequently, LysoTracker-Green DND-26 was

used to track lysosomes, and then the co-localization of intracellular RhoB-OVA250-264 and lysosomes

were analyzed by confocal scanning microscope.

Figure S4. OVA257-276 specific spots elicited by free OVA250-264 peptide with or without MPLA.

C57BL/6 mice were s.c injected with different dose of free OVA250-264 peptide (A and B) or the

mixture of PEG-PE micelle encapsulating MPLA (PEG-PE/MPLA) and different dose of free OVA250-

264 peptide (C and D). 6 days later, the draining inguinal LNs were isolated. The OVA257-264 specific

spots (per 3*105 lymphocyte) were measured using ELISPOT.

All the data are given as mean ± SEM; n =3/group;

Figure S5. Antigen specific IgG is generated in mice after inoculation of PEG-PE micelle-based

vaccine

C57BL/6 mice were s.c injected with PEG-PE micelle-based vaccine encapsulating HPV E743-62 or

E71-60 peptides on day 0 and 14. On day 28, mice were sacrificed and serum was obtained. E743-62 and

E71-60 peptide specific IgG was measured using ELISA.

(A) Immunization schedule of vaccine

(B) E743-62 peptide specific IgG was measured after immunization with PEG-PE micelle-based vaccine

encapsulating E743-62 peptide. The group of PEG-PE micelle encapsulating E743-62 peptide and free

E743-62 peptide was control.

(C) E71-60 peptide specific IgG was measured after immunization with PEG-PE micelle-based vaccine

encapsulating E71-60 peptide. The group of PEG-PE micelle encapsulating E71-60 peptide and free

E71-60 peptide was control.

All the data are given as mean ± SEM; n =3/group; *P<0.05; **P<0.01; ***P<0.001;