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. 2021 Apr 1;11(10):3134–3149. doi: 10.1016/j.apsb.2021.03.039

Figure 1.

Figure 1

TBM-1 decreases PD-L1 abundance effectively in tumor cells. (A) PD-L1 and MHC-I levels in H157 cells treatment with 5 μmol/L of 19 selected compounds for 36 h were analyzed by flow cytometry. Results are shown as fold change by comparing mean fluorescence intensity (MFI) of PD-L1 and MHC-I to vehicle-treated cells. TBM-1, tubeimoside-1; Cbf, cinobufagin. (B) Chemical structure of TBM-1. (C) Cell death effect of TBM-1 (5 μmol/L) and cinobufagin (5 μmol/L) on H157 cells were determined by cell impedance assay. (D) Flow cytometry analyzing the membrane PD-L1 levels in H157 and A375 cells treatment with TBM-1 (5 μmol/L) for 24 h. Statistic of MFI of PD-L1 is shown at the right. (E) H460, H157, A375 and A2058 cells were treated with indicated concentrations of TBM-1 for 24 h, or treated with 5 μmol/L of TBM-1 for the indicated times, the PD-L1 levels were detected by immunoblotting. Quantifications of PD-L1 to GAPDH were shown at the right. (F) Cellular PD-L1 and IDO1 levels in A549 or H1299 cells pretreatment with TBM-1 (5 μmol/L, 2 h) and IFN-γ (5 ng/mL) stimulation for 24 h without removing TBM-1 were determined by immunoblotting. Quantifications of PD-L1 to GAPDH were shown on the bottom. (G) Membrane PD-L1 levels in A549 or H1299 cells pretreatment with TBM-1 (5 μmol/L, 2 h) and IFN-γ (5 ng/mL) stimulation for 24 h without removing TBM-1 were detected by flow cytometry. Statistic MFI of PD-L1 is shown. Data are presented as mean ± SD, n = 3; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ns, no significant.