Fig. 5

P5C antibody counter the effect of P5C in vitro and in vivo. A Human primary CD3⁺ T cells were pretreated with P5C and P5C Ab then stimulated for 3 days with anti-CD3/CD28 beads. Shown is the percentage of cell proliferation by flow cytometry. The left side of graph is the representative result by flow cytometry. B Jurkat cells were pretreated with P5C and P5C Ab for 24 h. Western bolt showing the protein expression of SHP1 in Jurkat cells. An antibody to GAPDH was used as a loading control. C Jurkat cells were treated with P5C and P5C Ab, and ECAR assays were performed using the Seahorse XF24 analyzer. D Jurkat cells were pretreated with P5C and P5C Ab for 24 h. Western bolt showing the protein expression of PKM2 and p-PKM2 in Jurkat cells. An antibody to β-actin was used as a loading control. E Mice were inoculated subcutaneously with 1 × 10⁶ RM-1 cells to construct animal model, and treated with P5C and P5C Ab. The picture shows the tumors after harvesting. The line graph below shows the mean of tumor volume measured at the indicated number of days after mice were treated. F The infiltration of CD3⁺ T cells in tumor tissue detected by IF. The red light marked T cells. G IF showing SHP expression in CD3⁺ T cells. DAPI was used for marking nuclear. The red light marked T cells and green light marked SHP1. Error bars are SEM of biological replicates and ^*, ^#p < 0.05; ^***p < 0.01