Fig. 4: Knockdown of LCN2 alleviates HP-induced VSMCs calcification. | Cell Death & Disease

Fig. 4: Knockdown of LCN2 alleviates HP-induced VSMCs calcification.

From: Lipocalin-2 promotes CKD vascular calcification by aggravating VSMCs ferroptosis through NCOA4/FTH1-mediated ferritinophagy

Fig. 4

A–C VSMCs were transfected with LCN2 siRNA (siLCN2) or NC siRNA (siNC). The efficacy of transfection was verified by Western blot (n = 4 per group) and qRT-PCR (n = 3 per group). D Calcium deposition in VSMCs was assessed by Alizarin red staining (positive staining: red; scale bar: 200 μm) and quantified using hexadecylpyridinium chloride at an OD of 560 nm (E) (n = 6 per group). F Calcium deposition in VSMCs was assessed by calcium concentration assay and was normalised to the protein concentration (n = 3 per group). G Western blot analysis and quantification of the RUNX2 (H) and BMP2 (I) protein expression in siNC or siLCN2-transfected VSMCs exposed to HP (3.0 mM) or growth medium (CTL) (n = 3 per group). Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. qRT-PCR quantitative real-time polymerase chain reaction, HP high phosphate, VSMC vascular smooth muscle cell, siRNA small interfering RNA, NC negative control, OD optical density, CTL control, RUNX2 runt-related transcription factor 2, BMP2 bone morphogenetic protein 2, SD standard deviation.

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