Journal: Biomolecules

Article Title: ACSL4 Drives C5a/C5aR1–Calcium-Induced Fibroblast-to-Myofibroblast Transition in a Bleomycin-Induced Mouse Model of Pulmonary Fibrosis

doi: 10.3390/biom15081106

Figure Lengend Snippet: Pharmacological inhibition of ACSL4 alleviated pulmonary inflammation and fibrosis in the early fibrotic stage: ( A ) Scheme for BLM-induced lung fibrosis. The mice were treated with PRGL493 (250 μg/kg) or vehicle 1 h before BLM exposure on day 0, and then once a day. Mice were euthanized on day 7 after BLM modeling. ( B ) Weight changes of the mice; n = 8 for each group. ( C ) Lung coefficients of the mice; n = 8 for each group. ( D ) Hematoxylin–eosin (H&E) staining of lung tissues. Representative images are shown. Original magnification: 200×. ( E ) The number of nuclei per high-power field of lung tissues; n = 8 for each group. ( F ) Inflammatory index of the alveoli; n = 8 for each group. ( G ) The Ashcroft score; n = 8 for each group. ( H ) The expression of Acta2 , Col1a1 , Col3 , Tgf-β1 , tissue inhibitors of metalloproteinases-1 ( Timp1 ), and matrix metalloproteinases-2 ( Mmp-2 ) in lung tissues was quantified by qPCR; n = 8 for each group. ( I ) The expression of αSMA in the lung tissues was measured by WB. Representative bands and statistical plots are shown; n = 6 for each group. ( J ) Immunohistochemical staining of αSMA in lung tissues. Representative images and relative IOD analyses are shown. Original magnification: 200×; n = 8 for each group. ( K ) Immunohistochemical staining of collagen I in the lung tissues. Representative images and relative IOD analyses are shown. Original magnification: 200×; n = 8 for each group. ( L ) Masson staining of the lung tissues. Representative images and collagen volume fraction analyses are shown. Original magnification: 200×; n = 8 for each group. Data are the mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA followed by adjustments for multiple comparisons in panel ( B ), or Student’s t -test in panels ( C , E – L ); ns = not significant.

Article Snippet: Staining was performed according to the instructions of the Masson staining kit (Servicebio, Wuhan, China, Cat#: G1006).

Techniques: Inhibition, Staining, Expressing, Immunohistochemical staining

Journal: Biomolecules

Article Title: ACSL4 Drives C5a/C5aR1–Calcium-Induced Fibroblast-to-Myofibroblast Transition in a Bleomycin-Induced Mouse Model of Pulmonary Fibrosis

doi: 10.3390/biom15081106

Figure Lengend Snippet: Pharmacological inhibition of ACSL4 alleviated pulmonary inflammation and fibrosis in the chronic fibrotic stage. ( A ) Scheme for BLM-induced lung fibrosis. The mice were treated with PRGL493 (250 μg/kg) or vehicle 1h before BLM exposure on day 0, and then once a day. Mice were euthanized on day 21 after BLM modeling. ( B ) Weight changes of the mice; n = 6 for each group. ( C ) Lung coefficients of the mice; n = 6 for each group. ( D ) H&E staining of the lung tissues. Representative images are shown. Original magnification: 200×. ( E ) Number of nuclei per high-power field of the lung tissues; n = 6 for each group. ( F ) Inflammatory index of the alveoli; n = 6 for each group. ( G ) The Ashcroft score; n = 6 for each group. ( H ) The expression of Acta2 , Col1a1 , Col3 , Tgf-β1 , Timp-1 , and Mmp-2 in lung tissues was quantified by qPCR; n = 5 for each group. ( I ) WB analysis of αSMA in lung tissues. Representative bands and statistical plots are shown; n = 5 for each group. ( J ) Immunohistochemical staining of αSMA in lung tissues. Representative images and relative IOD analyses are shown. Original magnification: 200×; n = 6 for each group. ( K ) Immunohistochemical staining of collagen I in lung tissues. Representative images and relative IOD analyses are shown. Original magnification: 200×; n = 6 for each group. ( L ) The Masson staining of the lung tissues. Representative images and collagen volume fraction analyses are shown. Original magnification: 200×; n = 6 for each group. Data are the mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA followed by adjustments for multiple comparisons in panel ( B ), or Student’s t -test in panels ( C , E – L ). ns = not significant.

Article Snippet: Staining was performed according to the instructions of the Masson staining kit (Servicebio, Wuhan, China, Cat#: G1006).

Techniques: Inhibition, Staining, Expressing, Immunohistochemical staining

Journal: Biomolecules

Article Title: ACSL4 Drives C5a/C5aR1–Calcium-Induced Fibroblast-to-Myofibroblast Transition in a Bleomycin-Induced Mouse Model of Pulmonary Fibrosis

doi: 10.3390/biom15081106

Figure Lengend Snippet: Effects of blocking the C5a/C5aR1 signaling on the early stage of lung fibrosis: ( A ) Scheme for the BLM-induced lung fibrosis. The mice were treated with PMX53 (1 mg/kg) or vehicle 1 h before BLM exposure on day 0, and then once a day. Mice were euthanized on day 7 after BLM modeling. ( B ) Weight changes of the mice; n = 8 for each group. ( C ) Lung coefficients of the mice; n = 8 for each group. ( D ) H&E staining of the lung tissues. Representative images are shown. Original magnification: 200×. ( E ) The number of nuclei per high-power field of lung tissues; n = 8 for each group. ( F ) The inflammatory index of the alveoli; n = 8 for each group. ( G ) The Ashcroft score; n = 8 for each group. ( H , I ) The expression of Acta2 , Col1a1 , Col3 , and Fn1 in lung tissues was quantified by qPCR; n = 6 for each group. ( J ) The expression of αSMA protein in lung tissues. Representative immunohistochemical plots and statistical plots are shown. Original magnification: 200×; n = 8 for each group. ( K , L ) Immunohistochemical staining of collagen I in lung tissues. Representative images and relative IOD analyses are shown. Original magnification: 200×; n = 8 for each group. ( M ) Masson staining of the lung tissues. Representative images and collagen volume fraction analyses are shown. Original magnification: 200×; n = 8 for each group. ( N , O ) WB analysis of the ACSL4 and αSMA protein in lung tissues. Representative bands and statistical plots are shown; n = 6 for each group. ( P ) The expression of Acsl4 in lung tissues was quantified by qPCR; n = 6 for each group. ( Q ) Immunohistochemical staining of ACSL4 protein in the lung tissues. Representative images and relative IOD analyses are shown; n = 8 for each group. Data are the mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA followed by adjustments for multiple comparisons in panel ( B ), or Student’s t -test in panels ( C , E – J , L , M , O – Q ); ns = not significant.

Article Snippet: Staining was performed according to the instructions of the Masson staining kit (Servicebio, Wuhan, China, Cat#: G1006).

Techniques: Blocking Assay, Staining, Expressing, Immunohistochemical staining

Journal: Biomolecules

Article Title: ACSL4 Drives C5a/C5aR1–Calcium-Induced Fibroblast-to-Myofibroblast Transition in a Bleomycin-Induced Mouse Model of Pulmonary Fibrosis

doi: 10.3390/biom15081106

Figure Lengend Snippet: Inhibition of C5a/C5aR1 signaling reduced ACSL4 levels and attenuated lung inflammation and fibrosis in the chronic stage of pulmonary fibrosis: ( A ) Scheme of BLM-induced lung fibrosis. Mice were euthanized on day 21 after BLM modeling. ( B ) Weight changes of the mice; n = 6 for each group. ( C ) Lung coefficients of the mice; n = 6 for each group. ( D ) H&E staining of the lung tissues. Representative images are shown. Original magnification: 200×. ( E ) Number of nuclei per high-power field of the lung tissues; n = 6 for each group. ( F ) The inflammatory index of the alveoli; n = 6 for each group. ( G ) The Ashcroft score; n = 6 for each group. ( H , I ) The expression of Acta2 , Col1a1 , Col3 , and Fn1 in lung tissues was quantified by qPCR; n = 5 for each group. ( J ) Immunohistochemical staining of αSMA in lung tissues. Representative images and relative IOD analyses are shown. Original magnification: 200×; n = 6 for each group. ( K , L ) Immunohistochemical staining of collagen I in lung tissues. Representative images and relative IOD analyses are shown. Original magnification: 200×; n = 6 for each group. ( M ) Masson staining of lung tissues. Representative images and collagen volume fraction analyses are shown. Original magnification: 200×; n = 6 for each group. ( N , O ) WB analysis of the ACSL4 and αSMA proteins in lung tissues. Representative bands and statistical plots are shown; n = 6 for each group. ( P ) The expression of Acsl4 in lung tissues was quantified by qPCR; n = 6 for each group. ( Q ) Immunohistochemical staining of ACSL4 in lung tissues. Representative images and relative IOD analyses are shown; n = 6 for each group. Data are the mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA followed by adjustments for multiple comparisons in panel ( B ), or Student’s t -test in panels ( C , E – J , L , M , O – Q ); ns = not significant.

Article Snippet: Staining was performed according to the instructions of the Masson staining kit (Servicebio, Wuhan, China, Cat#: G1006).

Techniques: Inhibition, Staining, Expressing, Immunohistochemical staining