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Fig. 6 | Respiratory Research

Fig. 6

From: Down-regulation of USP13 mediates phenotype transformation of fibroblasts in idiopathic pulmonary fibrosis

Fig. 6

Levels of PTEN ubiquitination and degradation in USP13-deficient lung fibroblasts. a Endogenous USP13 was immunoprecipitated from MRC-5 whole cell lysates and immunoblotted with antibodies against USP13 and PTEN (left panel), while endogenous PTEN was immunoprecipitated from MRC-5 whole cell lysates and immunoblotted with antibodies against PTEN and USP13 (right panel). b Co-localization of USP13 and PTEN. PTEN (red) and USP13 (green) were labeled by immunofluorescence staining of MRC-5 cells. The right panel shows the overlay (merge) of PTEN, USP13, and nuclear DAPI staining (blue) in the same field to determine co-localization. Scale bar, 20 μm. c At 42 h after transfection with USP13 siRNA or NC siRNA, we added MG-132 (10 μM) for 6 h and harvested whole cell lysates. PTEN and ubiquitinated PTEN (Ub-PTEN) were assessed by immunoprecipitation and western blot. β-actin was used as a loading control. d Immunofluorescence staining of PTEN (red) and USP13 (green) in MRC-5 cells transfected with USP13 siRNA (USP13si) or negative control siRNA (NCsi) for 48 h. Nuclei were visualized by DAPI (blue) staining. Scale bar, 20 μm. (a–d) Data are representative of at least three independent experiments. e Representative serial lung sections from one IPF patient were stained for (a) H&E, (b) α-SMA, (c) PTEN, and (d) USP13. Images are representative of staining performed on samples from 11 individuals with IPF. Scale bar, 100 μm

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