Fig. 2 | Signal Transduction and Targeted Therapy

Fig. 2

From: Colony-stimulating factor 3 as a key mediator in the progression of idiopathic pulmonary fibrosis: a novel therapeutic target

Fig. 2

CSF3 knockout attenuates pulmonary fibrosis progression. a Schematic illustration of the bleomycin (BLM) intratracheal injection (IT) animal model and the procedure used to induce lung fibrosis in C57BL/6 mice. b and c Representative Masson’s trichrome staining (b, c) and immunohistochemistry for α-SMA and COL1A1 (c) of lung tissue from CSF3+/+ and CSF3−/− mice after BLM or PBS treatment (n = 5 per group). Scale bar: 25 μm, Scale bar: 100 μm. d–i qRT-PCR analysis of α-SMA, COL1A1, and CSF3 expression (d, e), hydroxyproline content assay (f), qRT-PCR analysis of prolyl hydroxylases (P4HA1, P4HA2, P4HA3) (g), matrix metalloproteinase 2 (MMP2) (h) and tissue inhibitors of metalloproteinases (TIMP1/2) (i) expression in lung tissue from CSF3+/+ and CSF3−/− mice after BLM or PBS treatment. j qRT-PCR analysis of α-SMA, COL1A1, and CSF3 expression in human lung fibroblasts isolated from idiopathic pulmonary fibrosis patients (IPDF) and transfected with siRNA targeting CSF3 (si-CSF3). k Western blot analysis of CSF3, αSMA, COL1A1, and p-STAT3 in IPDF cells transfected with si-CSF3 in IPDF. β-Actin was used as a loading control. l Representative immunofluorescence images of α-SMA and COL1A1 in IPDF transfected with si-CSF3. Scale bars: 200 μm. m The spontaneous Matrigel-invading capacity of IPDF transfected with si-CSF3 or si-Control. Scale bars: 200 μm. Statistical significance was determined using ANOVA with multiple comparison or t-test

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