Fig 1: Optimization of p53-cAb-DNA (2.5, 5, 10, and 25 fmol/mL) and anti-human-IgG-FB (0.015, 0.035, 0.07, 0.175, and 0.35 fmol/mL) concentrations for PIC detection. The PIC (pg/mL) levels were (a) 5000, (b) 2500, (c) 1250, (d) 625, (e) 312; (f) linearity in the serial dilution of 5000 pg/mL of PIC using 0.07 fmol/mL of anti-human-IgG-FB and 10 fmol/mL of p53-cAb-DNA.
Fig 2: Optimization of p53-dAb-FB (0.015, 0.035, 0.07, 0.175, and 0.35 fmol/mL) and p53-cAb-DNA (2.5, 5, 10, and 25 fmol/mL) concentrations for Tp53 detection. The Tp53 (pg/mL) levels were (a) 5000, (b) 2500, (c) 1250, (d) 625, (e) 312; (f) linearity in the serial dilution of 5000 pg/mL of Tp53 using 0.07 fmol/mL of p53-dAb-FB and 10 fmol/mL of p53-cAb-DNA.
Fig 3: General procedure for detecting (a) Tp53 and (b) p53-anti-p53-autoantibody complex (PIC) in the plasma samples.
Fig 4: Standard curves for (a) Tp53 and (b) PIC detection.
Fig 5: Sb9 decreases GZMB expression by suppressing TP53 expression in CD8 T cells.a PCR and qRT-PCR results for the GZMB promoter in CD8 T cells following ChIP using TP53 antibody. Mean± SD, n = 3 biological replicates, data represent two independent experiments. b Representative image of EMSA using probes corresponding to the WT or MUT GZMB promoter together with TP53 protein and TP53 antibody. EMSA was repeated independently three times with similar results. c GZMB mRNA level in CD8 T cells transfected with or without TP53 shRNAs. Mean± SD, n = 3 biological replicates, data represent two independent experiments. d Transcription activity of GZMB analyzed by relative luciferase reporter activity assay in scrambled and TP53 shRNAs-transfected CD8 T cells. Mean± SD, n = 3 biological replicates, data represent two independent experiments. e TP53 mRNA level in CD8 T cells transfected with or without the Sb9 expression vector. f TP53 protein level in CD8 T cells transfected with or without Sb9 expression vector. Mean± SD, n = 3 biological replicates, data represent two independent experiments. Sb9 SerpinB9, GZMB granzyme B, WT wild type, MUT mutation, OE overexpression, EMSA, electrophoretic mobility shift assay, PCR polymerase chain reaction, qRT-PCR quantitative reverse transcription-PCR. All statistical tests were two-sided. Statistical significance was assessed using one-way ANOVA followed by post-hoc Tukey’s test except for (b). Adjustments were made for multiple comparisons. *P < 0.05; **P < 0.01; ****P < 0.0001. Source data are provided as a Source Data file.
Supplier Page from Abcam for Recombinant Human p53 protein