Journal: Journal of Cellular and Molecular Medicine

Article Title: Orderly Regulation of Macrophages and Fibroblasts by Axl in Bleomycin‐Induced Pulmonary Fibrosis in Mice

doi: 10.1111/jcmm.70321

Figure Lengend Snippet: BLM modelling in mice with Axl total inhibition or knockout. (A) After BLM treatment, mice were administered R428 or ONO‐7475 from Day 0 to Day 21. (B) Body weight curves of the aforementioned mice. Initially, control group ( n = 6), BLM group ( n = 7), R428 group ( n = 10) and ONO‐7475 group ( n = 10). Three mice in the R428 group, one in the ONO‐7475 group and one in the BLM group perished during the administration of BLM. # represents the death of one mouse. (C) H&E staining and Masson staining after sampling on Day 21. Scale bars: All 20 μm. Control group ( n = 6), BLM group ( n = 6), R428 group ( n = 7) and ONO‐7475 group ( n = 9). (D) Ashcroft scores of mice in Figure C. (E) Axl −/− or Axl +/− mice treated with BLM were sampled on Day 21 for H&E staining and Masson staining. Scale bars: All 20 μm. (F) The weight curves of the mice in Figure E. (G) Ashcroft scores of mice in Figure E. (H) Hydroxyproline contents in lung tissues of mice in Figure E. (I) Immunohistochemistry of paraffin sections of mice's lung tissues in Figure E. Scale bars: All 50 μm. (J, K) Immunohistochemical quantification of CD68 (J) and α‐SMA (K) protein levels in Figure I. n = 4–10 mice/group, **** p < 0.0001. # represents the death of a mouse. ## means two dead mice.

Article Snippet: Wild‐type inhibitor groups received intragastric administration of R428 [ , ] (5 mg/kg, MedChem Express, Monmouth Junction, NJ, USA), the inhibitor of Axl or ONO‐7475 [ ] (10 mg/kg, Selleck Chemicals, Houston, TX, USA), the inhibitor of both Axl and Mer, respectively, while the blank control group and the BLM group received the same dose of sodium carboxymethylcellulose (CMC‐Na, Solarbio, Beijing, China) solution.

Techniques: Inhibition, Knock-Out, Control, Staining, Sampling, Immunohistochemistry, Immunohistochemical staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Orderly Regulation of Macrophages and Fibroblasts by Axl in Bleomycin‐Induced Pulmonary Fibrosis in Mice

doi: 10.1111/jcmm.70321

Figure Lengend Snippet: Inhibition of Axl in the acute phase of pulmonary fibrosis. (A) After BLM administration, mice were given R428 and ONO‐7475, respectively, and the administration time was from Day 0 to Day 7. (B‐C) Ashcroft scores (B), H&E staining and Masson staining (C) after sampling on Day 21. Scale bars: All 20 μm. (D) Protein levels of collagen Type I and α‐SMA were determined by immunohistochemistry. Scale bars: All 50 μm. (E, F) Immunohistochemical quantification of collagen Type I (E) and α‐SMA (F) protein levels in mice in Figure D. n = 9–10 mice/group, */**/*** p < 0.05/0.01/0.001.

Article Snippet: Wild‐type inhibitor groups received intragastric administration of R428 [ , ] (5 mg/kg, MedChem Express, Monmouth Junction, NJ, USA), the inhibitor of Axl or ONO‐7475 [ ] (10 mg/kg, Selleck Chemicals, Houston, TX, USA), the inhibitor of both Axl and Mer, respectively, while the blank control group and the BLM group received the same dose of sodium carboxymethylcellulose (CMC‐Na, Solarbio, Beijing, China) solution.

Techniques: Inhibition, Staining, Sampling, Immunohistochemistry, Immunohistochemical staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Orderly Regulation of Macrophages and Fibroblasts by Axl in Bleomycin‐Induced Pulmonary Fibrosis in Mice

doi: 10.1111/jcmm.70321

Figure Lengend Snippet: Axl inhibition at the stage of acute inflammatory reaction can aggravate lung injury. (A) After BLM treatment, mice were given R428 or ONO‐7475 and the administration time was from Day 0 to Day 3/7 by intragastric administration, and samples were collected on Day 3/7. (B–D) H&E staining and lung injury scores were performed after sampling on Day 3 or 7. Scale bars: All 20 μm. (E–H) The expression levels of IL‐1β and TNF‐α were detected by ELISA after grinding mice's lung tissues on Day 3 or 7. (I) Immunohistochemistry of mice's lung tissues on Day 3 or 7. Scale bars: All 50 μm. (J–M) Protein levels of CD68 (J) and MPO (K) in lung tissues on Day 3 and CD68 (L) and MPO (M) in lung tissues on Day 7 of immunohistochemical quantification in mice in Figure I. n = 3–6 mice/group, */** p < 0.05/0.01.

Article Snippet: Wild‐type inhibitor groups received intragastric administration of R428 [ , ] (5 mg/kg, MedChem Express, Monmouth Junction, NJ, USA), the inhibitor of Axl or ONO‐7475 [ ] (10 mg/kg, Selleck Chemicals, Houston, TX, USA), the inhibitor of both Axl and Mer, respectively, while the blank control group and the BLM group received the same dose of sodium carboxymethylcellulose (CMC‐Na, Solarbio, Beijing, China) solution.

Techniques: Inhibition, Staining, Sampling, Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Immunohistochemical staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Orderly Regulation of Macrophages and Fibroblasts by Axl in Bleomycin‐Induced Pulmonary Fibrosis in Mice

doi: 10.1111/jcmm.70321

Figure Lengend Snippet: Inhibiting Axl during the fibrotic phase reduced pulmonary fibrosis. (A) After BLM treatment, mice were given R428 or ONO‐7475, and the administration time was from Day 7 to Day 21. (B) Body weight curves of mice within 21 days. (C) H&E staining and Masson staining were performed after paraffin sections of the above mice's lung tissues. Scale bars: All 20 μm. (D) Ashcroft scores of mice in Figure C. (E) Hydroxyproline quantifications in lung tissues on Day 21. (F) Lung tissues were lysed and WB was performed, Collagen Type I and α‐SMA results were obtained. Three samples in each group were mixed and repeated three times. (G, H) Quantitative results of collagen Type I (G) and α‐SMA (H) in Figure F. n = 3–10 mice/group. */**/***/**** p < 0.05/0.01/0.001/0.0001.

Article Snippet: Wild‐type inhibitor groups received intragastric administration of R428 [ , ] (5 mg/kg, MedChem Express, Monmouth Junction, NJ, USA), the inhibitor of Axl or ONO‐7475 [ ] (10 mg/kg, Selleck Chemicals, Houston, TX, USA), the inhibitor of both Axl and Mer, respectively, while the blank control group and the BLM group received the same dose of sodium carboxymethylcellulose (CMC‐Na, Solarbio, Beijing, China) solution.

Techniques: Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Orderly Regulation of Macrophages and Fibroblasts by Axl in Bleomycin‐Induced Pulmonary Fibrosis in Mice

doi: 10.1111/jcmm.70321

Figure Lengend Snippet: Inhibition of Axl or FAK can reduce the activation of PXN in fibroblasts. The extracted primary fibroblasts were treated with inhibitors for 4 h, then TGF‐β or PBS were added and follow‐up experiments were performed 48 h later. The concentration of R428 was 2 mmol/L, and the concentration of PF‐573228 was 3 mmol/L. (A) Photograph of the cell invasion experiment described above (original magnification: ×200). (B) The invasion experiment of the above cells was quantitative, and five random visual fields were selected from each Transwell under ×400 magnification for statistics. (C) Cell proteins were extracted and the protein levels of β‐tubulin, PXN, p‐PXN, FAK and p‐FAK were assessed by western blotting. (D) Protein quantification of p‐FAK in Figure C. (E) Protein quantification of p‐paxillin in Figure C. n = 3 mice/group, */*** p < 0.05/0.001.

Article Snippet: Wild‐type inhibitor groups received intragastric administration of R428 [ , ] (5 mg/kg, MedChem Express, Monmouth Junction, NJ, USA), the inhibitor of Axl or ONO‐7475 [ ] (10 mg/kg, Selleck Chemicals, Houston, TX, USA), the inhibitor of both Axl and Mer, respectively, while the blank control group and the BLM group received the same dose of sodium carboxymethylcellulose (CMC‐Na, Solarbio, Beijing, China) solution.

Techniques: Inhibition, Activation Assay, Concentration Assay, Western Blot