Fig 1: Pharmacokinetic analysis of CTLA4-FasL. CTLA4-FasL at different doses was injected s.c. to mice at a total volume of 150 µl per mouse. Mice were sacrificed at various time points (0-24 h) post injection. CTLA4-FasL level in plasma was quantified by Human Soluble CTLA-4 ELISA kit.
Fig 2: Schematic model of the CTLA4-FasL homo-hexamer. Monomer represents one domain of the CTLA-4 connected to one domain of the FasL. Trimer is composed of the FasL trimer and three domains of the CTLA-4, while two of them are connected by a disulfige bridge. Two CTLA4-FasL trimers may form a homo-hexamer, made of dimer of trimmers or trimer of dimers, resulting in a shift towards a small fraction of a dodecamer state.
Fig 3: CTLA4-FasL molecular structure. (A) SDS-PAGE analysis of the CTLA4-FasL under reducing conditions, coomassie G-250 stain. (B) Western blot analysis using anti CTLA4 antibody following enzymatic removal of the N-glycan chains from the protein. (C) Iso-Electric Focusing analysis of CTLA4-FasL at pH3-7 and pH3-10.
Fig 4: CTLA-4•FasL inhibits tumor growth in vivo . (A) Ex vivo JY cells were injected subcutaneously to irradiated NUDE mice. At day 5 after JY injection, tumor volume was calculated and mice were treated daily with subcutaneous injections of CTLA4-FasL (10 µg twice a day for 5 days for one treatment course or 2 µg twice a day for 4 days for four consecutive weeks). Control animals received similar volume of PBS. The results represent the mean +/- SE volume of tumors (n = 10 for each group). * - P <0.05, **P <0.01 between PBS group and treatment group (B) Five days post s.c. injection of human JY tumor cells to irradiated-NUDE mice the mice were treated with 10 ug CTLA4-FasL or PBS twice a day for 5 consecutive days. Tumors were harvested one hour after last injection, fixated and embedded in paraffin, and tissue sections were processed and stained with anti cleaved caspase 3 antibody. (C) 8 days following last CTLA4-FasL (100 µg a day, 4 days), or vehicle injection mice livers were harvested, fixated and embedded in paraffin, and stained with H&E.
Fig 5: CTLA4-FasL inhibits tumor growth in vivo . Ex vivo JY cells were injected subcutaneously to irradiated NUDE mice. At day 5 after JY injection, tumor volume was calculated and mice were treated daily with subcutaneous injections of CTLA4-FasL or PBS for 5 days. (A) Tumor volume. Results represent the mean +/- SE volume of tumors, (n = 10 for each group). *P < 0.05 between PBS group and 10 ug*2 or PBS vs 25 ug*2. **P <0.01 respectively. (B) The survival curve of the experimental groups and control.
Supplier Page from BioLegend for LEGEND MAX(TM) Human Soluble CTLA-4 ELISA Kit