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. 2021 Sep 6;12:5296. doi: 10.1038/s41467-021-25673-6

Fig. 3. LRP1 depletion induces OCN through increasing FoxO nuclear export in ECs.

Fig. 3

a, b LRP1 was associated with FoxOs. Lysates of HEK 293 cells containing stably expressed Flag-LRP1β (a) or MLECs (b) were immunoprecipitated with anti-Flag or anti-LRP1 resin and blotted for FoxOs. c–e LRP1 depletion in MLECs led to FoxO1 nuclear export. MLECs were transfected with LRP1 or control siRNAs and subjected for immunofluorescence imaging (b) or subcellular fractionation assays (d, e). MLECs were stained for FoxO1 (green) and the nucleus (DAPI, blue, b) and the intensity ratio of FoxO1 signals in the nucleus compared to that in cytosol was quantified. Scale bar, 10 μm. TCL, total cell lysates. f Constitutively active FoxO1 (CA-FoxO1) inhibited LRP1 depletion-induced OCN, analyzed with real-time PCR. n = 5 (c), 3 (e), and 4 (f). NS, not significant. Data are presented as mean ± SEM. Analysis was two-way ANOVA followed by Fisher’s LSD multiple comparison test (f) or unpaired two-tailed Student’s t-test (c, e).