The three plasmid systems, including target gene plasmid (pAAV-CAG-mCherry), capsid plasmid (pAAV-6, pAAV-1, pAAV-7m8, pAAV-7, pAAV-8, pAAV-10) and helper plasmid (pHelper), were all purchased from Fenghui Biotechnology (Hunan, China). The complementary DNA (cDNA) encoding the mouse PTEN gene was synthesized from GENEWIZ (Suzhou, China). pAAV-CAG-PTEN was obtained by replacing mCherry gene in pAAV-CAG-mCherry with PTEN gene. Reverse the PTEN gene sequence in pAAV-CAG-PTEN to get pAAV-CAG-PTENR.
The anti-PD-1 used in this study is AUNP-12, a polypeptide purchased from Selleck (USA) with a purity of 99.20%. Cy5-AUNP-12 was purchased from Guoping pharmaceutical co (Anhui, China) with a purity of 90%. CpG was synthesized in GENEWIZ (Suzhou, China) with 90% purity.
Antibodies for Western blot (WB) and Immunofluorescence (IF): anti-mouse PTEN, CRT, β-actin and anti-rabbit FITC, CY3 fluorescent secondary antibodies were purchased from ABclonal Tech (Wuhan, China). Antibodies for flow cytometry test: CD11c-PE, CD80-FITC, CD86-APC, CD3-APC, CD8-FITC, CD4-APC-eflour780, CD69-PE, CD274-PE, CD11b-FITC, CD206-PE-Cy7, CD86-APC, IL-4-PE, IFN-γ-APC, TNF-α-eflour450, IL-2-PE5.5, CD4-APC-eflour780, CD44-APC, CD62L-PE were all purchased from Invitrogen (USA).
The anti-PD-1 used in this study is AUNP-12, a polypeptide purchased from Selleck (USA) with a purity of 99.20%. Cy5-AUNP-12 was purchased from Guoping pharmaceutical co (Anhui, China) with a purity of 90%. CpG was synthesized in GENEWIZ (Suzhou, China) with 90% purity.
Antibodies for Western blot (WB) and Immunofluorescence (IF): anti-mouse PTEN, CRT, β-actin and anti-rabbit FITC, CY3 fluorescent secondary antibodies were purchased from ABclonal Tech (Wuhan, China). Antibodies for flow cytometry test: CD11c-PE, CD80-FITC, CD86-APC, CD3-APC, CD8-FITC, CD4-APC-eflour780, CD69-PE, CD274-PE, CD11b-FITC, CD206-PE-Cy7, CD86-APC, IL-4-PE, IFN-γ-APC, TNF-α-eflour450, IL-2-PE5.5, CD4-APC-eflour780, CD44-APC, CD62L-PE were all purchased from Invitrogen (USA).