Fig. 4: KIM-1 mediates bHDL uptake by injured RTECs. | Nature Communications

Fig. 4: KIM-1 mediates bHDL uptake by injured RTECs.

From: High-density lipoprotein nanoparticles spontaneously target to damaged renal tubules and alleviate renal fibrosis by remodeling the fibrotic niches

Fig. 4

a Uptake of bHDL under 4°C and with different endocytic inhibitors. Data are mean ± SD (n = 3 independent samples, two-tailed unpaired t-test). b Effect of D-4F on bHDL uptake in HK-2 cell. Data are mean ± SD (n = 3 independent samples, one-way ANOVA with Dunnett’s multiple comparisons test and adjustment applied). c Uptake of DiD-bHDL in normal and cisplatin-injured HK-2 cells for 2 hours, with or without pretreatment using KIM-1 antibody (30 µg/ml, 0.5 hours). Data are mean ± SD (n = 3 independent samples, two-tailed unpaired t-test). The control refers to untreated cells; KIM-1 antibody pretreated refers to cells with antibody pretreatment. d Representative confocal images of cisplatin-injured HK-2 cells after 2 hours of exposure. The nuclei were stained DAPI (blue), KIM-1 was labeled in green and DiD-bHDL was visualized in red. Scale bars, 20 µm. e Representative immunofluorescence images of colocalization of KIM-1(green) with DiD-bHDL or DiD-liposome (red) in the kidney. Nuclei were stained DAPI (blue). Scale bars, 50 µm. f Quantification of the colocalization of KIM-1 and DiD-bHDL or DiD-liposome by Pearson’s correlation coefficient. The Pearson’s correlation coefficient was measured by Colocalization Finder with image J. Data are mean ± SD (n = 3 biologically independent samples, two-tailed unpaired t-test). g, h Uptake of DiD-bHDL in HK-2 cells with KIM-1 protein knockdown(g) or overexpression(h) for 2 hours after 24-hour cisplatin treatment(2 µg/ml). Data are mean ± SD (n = 3 biologically independent samples, two-tailed unpaired t-test). i Western blot analysis of SR-BI expression in normal and cisplatin-injured HK-2 cells treated with cisplatin(2 µg/ml) for 24 hours. n = 3 independent samples. j Semi-quantification of SR-BI protein level (i) using image J, with β-actin as the internal control. Predicted protein weights: β-actin (43 kDa) and SR-BI (74 kDa). Data are mean ± SD (n = 3 biologically independent samples, two-tailed unpaired t-test). DiD-liposome DiD-laden liposomes, DiD-bHDL DiD-loaded biomimetic high-density lipoprotein nanoparticles, control untreated cells, CDDP-induced cisplatin-induced injured cells, KIM-1-/- HK-2 HK-2 cells with HAVCR1 knocked down, OE-KIM-1 HK-2 HK-2 cells with HAVCR1 overexpression. Source data are listed in the Source Data file.

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