HMGB1 Antibody - #AF7020

Figure 3. Effects of TPP-TAM@Alb on the expression of PD-L1 and TGF-β proteins in 4T1 cells. (A, B) The typical image of PD-L1 and TGF-β proteins detected by Western blot assay after different treatments. (C, D) Quantification of the expression levels of PD-L1 and TGF-β proteins after various treatments (n = 3). (E) The typical image of AMPK and pAMPK proteins detected by Western blot assay after TPP-TAM@Alb treatment in the presence or absence of Com C (used as AMPK inhibitor to depress AMPK pathway). (F, G) Evaluation and quantification of the expression levels of PD-L1 proteins by Western blot assay after TPP-TAM@Alb treatment in the presence or absence of Com C (n = 3). (H) The representative immunofluorescence images of CRT and HMGB1 after PTX@Alb, TPP-TAM@Alb, or PTX@Alb and TPP-TAM@Alb combination therapy, scale bar = 10 μm. All data are presented as mean ± SD. ∗P < 0.05, ∗∗P < 0.01. ns, not significant, P > 0.05.

Figure 3. Uptake and efficacy of sHDLs. (A) Flow cytometry analysis of time-dependent cellular uptake of Cy5-labelled sHDL by 4T1 cells. (B) Confocal images of 4T1 cells treated with Cy5-labelled sHDL at different time points. The nucleus and β-actin were stained with DAPI and Actin-Tracker Green, respectively. Scale bar = 20 μm. (C) Cell viability of 4T1 cells after 48 h exposure to LEN, L-sHDL, and LV-sHDL of various concentrations. Quantification of extracellular ATP (D) and HMGB1 (E) in the medium of 4T1 cells after the treatment of different sHDLs. (F) Confocal images of 4T1 cells after 4 h treatment with different sHDLs and another 12 h incubation with medium. The nucleus, HMGB1, and CRT were stained with DAPI, anti-HMGB1 antibody-Alexa Fluor® 647, and anti-CRT antibody-Alexa Fluor® 488, respectively. Scale bar = 20 μm. (G) Quantification of IFN-α and IFN-β secreted by 4T1 cells after the treatment of different sHDLs. Statistical significance was calculated using a two-sided one-way ANOVA test. The data are presented as the mean ± SD (n = 3).