Journal: Molecular Medicine Reports

Article Title: Uremic toxin p-cresyl sulfate enhances the calcification of aortic valvular interstitial cells via klotho/sirtuin-1 signaling

doi: 10.3892/mmr.2026.13872

Figure Lengend Snippet: Effect of PCS on klotho and HIF-1α expression. Western blotting and semi-quantification of (A) klotho and (B) HIF-1α protein expression in control and PCS-treated valvular interstitial cells. PCS-treated cells were incubated with PCS (100 µM) for 7 days (n=5). β-actin was used as an internal control. *P<0.05. PCS, p-cresol; HIF-1α, hypoxia-inducible factor-1α.

Article Snippet: The membranes were blocked with 10% non-fat milk in PBS for 1 h at room temperature and incubated overnight at 4°C with primary antibodies against RUNX2 (1:2,000 dilution; cat. no. 12556; Cell Signaling Technology), acetylated-NF-κB (1:2,000 dilution; acetyl-NF-κB; cat. no. 3045; Cell Signaling Technology, Inc.), NF-κB (1:3,000 dilution; cat. no. 8242; Cell Signaling Technology, Inc.), HIF-1α (1:1,000 dilution; cat. no. NB100-479; Novus Biologicals), klotho (1:1,000 dilution; cat. no. LS-C145689; LifeSpan BioSciences, Inc.) and β-actin (1:6,600 dilution; cat. no. ab6276; Abcam).

Techniques: Expressing, Western Blot, Control, Incubation

Journal: Molecular Medicine Reports

Article Title: Uremic toxin p-cresyl sulfate enhances the calcification of aortic valvular interstitial cells via klotho/sirtuin-1 signaling

doi: 10.3892/mmr.2026.13872

Figure Lengend Snippet: Effect of HIF-1α inhibition on PCS-induced VIC calcification. The upper panel depicts representative images of Alizarin Red S-stained VICs that have been incubated with or without the HIF-1α inhibitor PX-478 (0.5 µM) and with or without PCS (100 µM). VIC calcification was quantified by determining the total area of positive staining (red) per field (3 fields were observed per treatment group), as shown in the lower panel (n=3). Scale bar, 25 µm. *P<0.05. PCS, p-cresol; VIC, valvular interstitial cell; HIF-1α, hypoxia-inducible factor-1α.

Article Snippet: The membranes were blocked with 10% non-fat milk in PBS for 1 h at room temperature and incubated overnight at 4°C with primary antibodies against RUNX2 (1:2,000 dilution; cat. no. 12556; Cell Signaling Technology), acetylated-NF-κB (1:2,000 dilution; acetyl-NF-κB; cat. no. 3045; Cell Signaling Technology, Inc.), NF-κB (1:3,000 dilution; cat. no. 8242; Cell Signaling Technology, Inc.), HIF-1α (1:1,000 dilution; cat. no. NB100-479; Novus Biologicals), klotho (1:1,000 dilution; cat. no. LS-C145689; LifeSpan BioSciences, Inc.) and β-actin (1:6,600 dilution; cat. no. ab6276; Abcam).

Techniques: Inhibition, Staining, Incubation