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Fig. 4 | Respiratory Research

Fig. 4

From: FK506-binding protein 10 (FKBP10) regulates lung fibroblast migration via collagen VI synthesis

Fig. 4

FKBP10 interacts with COL6A1 and FBLN1 and regulates their expression. a, e Western Blot analysis of phLF treated with scrambled siRNA as control (sc) or FKBP10 siRNA (kd) and 2- phosphoascorbate (0.1 mM) in absence and presence of TGF-β1 (2 ng/mL) for 48h. Densitometric analysis and representative blots showing the effect of FKBP10 kd on the expression of Collagen VI alpha1 (COL6A1) (a) and fibulin-1 (FBLN1) (e) relative to β-actin as loading control (ACTB). b, c, d, f Quantitative reverse transcriptase-polymerase chain reaction analysis of phLF treated with sc siRNA as control or FKBP10 siRNA and 2-phosphoascorbate (0.1 mM) in absence and presence of TGF-β1 (2 ng/mL) for 48h. Transcript levels are shown as -ΔCt values of showing the effect of FKBP10 kd on collagen VI alpha 1 (COL6A1) (b), collagen VI alpha 2 (COL6A2) (c), collagen VI alpha 3 (COL6A3) (d) and fibulin-1C (FBLN1C) (f). DEAH (Asp-Glu-Ala-His) Box Polypeptide 8 (DHX8) was used as endogenous control. All data is based on eight (protein) or seven (mRNA) independent experiments. Statistical significance is indicated by horizontal brackets and asterisks. g-i Immunofluorescence staining of FKBP10 (green) and COL6A1 (red) (g), COL6A3 (red) (h) and FBLN1 (red) (i). 4`, 6-diamidino-2-phenylindole (DAPI) staining is shown in blue. The region of interest is indicated by a white square and enlarged in the picture below. White arrows specify examples for co-localization of FKBP10 with COL6A1, COL6A3 or FBLN1. Stainings were taken as z-stack and the enlarged pictures show one focal plane from this z-stack. Representative images were selected from three independent experiments. j Representative images of in situ localization of the interaction of FKBP10 and COL6A1, COL6A3, FBLN1, mouse IgG1κ (negative control), Golgin97 (negative control) and ERp57 (positive control), assessed by proximity ligation assay. Representative images were selected from three independent experiments, except for IgG negative control (n=1)

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