Fig 1: RCD profiling and PSGL-1 levels in hepatocytes from TAA-treated mice. (A) The RCD profile in ASGR1+ hepatocytes was assessed following TAA treatment using flow cytometry to evaluate changes in RCD levels. (B) Surface expression levels of PSGL-1 were measured in RCD-marker+ASGR1+ double-positive hepatocytes by flow cytometry. For example, in the apoptosis group, PSGL-1 levels were determined by identifying PSGL-1+ cells within the active caspase-3+ASGR1+ double-positive population. The “overall” groups were analyzed for PSGL-1 expression in ASGR1+ hepatocytes, independent of RCD markers. Vehicle control groups were normalized to a 1-fold change. ** p < 0.01 vs. respective vehicle-treated groups. The study was conducted with three independent experiments, each including two replicates per group [N = 3 (3 independent experiments), with n = 2 (2 technical replicates per experiment)]. A total of 12 mice were used in the study.
Fig 2: P-selectin-conjugated liposomes exert a rescue effect that is associated with the suppression of caspase-3 activities in mice. (A) Elevated surface PSGL-1 expression and (B) activated caspase-3 levels in TAA-treated mouse (wild type and Selplg−/−) hepatocytes in vivo were markedly rescued by treatments with P-selectin-conjugated liposomes (P-sel groups) loaded with caspase-3 inhibitor z-DEVD (5 × 107 liposomes, containing 2 μM z-DEVD/mouse). The measurements of untreated groups were normalized to 1-fold. ** p < 0.01, indicated markedly exacerbated vs. respective untreated control groups; # p < 0.05 indicated markedly ameliorated vs. respective IgG-Fc control groups. Three experiments with two mice per group (n = 6) were performed. A total of 48 (24 WT, 24 Selplg−/−) mice were used in the study.
Fig 3: Klotho mitigated IS-induced apoptotic responses. HK-2 cells were exposed to 500 μM IS for 24 h. Representative Western blot images and relative densitometric bar graphs of Bcl-2/b-actin, Bax/b-actin, and cytosolic cytochrome c/b-actin (A–D) in HK-2 cells stimulated to 500 mM IS for 24 h are shown. The activity of caspase 3 was assessed using flow cytometry (E). Annexin V was conducted for apoptosis assay (F). Results are expressed as means ± SD of three independent experiments. (* indicating p < 0.05 compared with the control group; # indicating p < 0.05 compared to IS-stimulated cells).
Supplier Page from Abcam for Cleaved Caspase-3 Staining Kit (Red)