Repression of allo-cell transplant rejection through CIITA ribonuclease P+ hepatocyte

Figure 1 Cleavage of CIITA substrate by M1-GS RNA in vitro (A): The mould plate RNA of CIITA pGM-3176 (3176-3560) include the cutting point of only M1-3408-GS, not M1-452-GS. (B): Schematic representation of targeting the CIITA mRNA by M1-RNA: GS encoding 12 or 11 nucleotides complemen-tary to CIITA was covalently linked to the 3’ end of M1-452-GS or M1-3408-GS. So M1-452-GS and M1-3408-GS specifily cut CIITA on the site of 452 and 3408 respectivly. (C): Sequence-specific cleavage of pGM-3176 substrate by M1-3408-GS: auto-radiograph of transcripts of M1-452-GSC (lane 1), M1-3408-GS (lane 2) and pGM-3176 (lane 3), and pGM-3176 substrate was incubated either with M1-3408-GS (lane 4), or with M1-452-GS (lane 5). So only M1-3408-GS (not M1-452-GS) could cleave pGM-3176.

Figure 2 Comparison of CIITA, MHCII mRNA abundance of psNAV-M1-3408-GS⁺ hepatocyte with void vector⁺ following IFN-γ induction The total RNA of psNAV-M1-3408-GS⁺ (p3408-H) or void vector⁺ (p-H) hepatocyte after IFN-γ (30 ng/ml, 3 d) induction was both extracted, then RT-PCR. 1: HLA-DR (335 bp); 2: HLA-DP (197 bp); 3: HLA-DQ (153 bp); 4: Ii (714 bp); 5: β-actin (310 bp); 6: CIITA (410 bp); 7: 100 bp DNA Ladder.

Figure 3 The secretion of IL-2 mRNA from PBMNC through RT-PCR. M: 100 bp DNA ladder; 1: Negative control; 2: The IL-2 from PBMNC stimulated by psNAV-M1-3408-GS⁺ hepato-cyte after IFN-γ induction; 3: The IL-2 from PBMNC stimu-lated by void vector control after IFN-γ induction.