Additional file 1 of IL-27 induces autophagy through regulation of the DNMT1/lncRNA MEG3/ERK/p38 axis to reduce pulmonary fibrosis
Creators: name: Ting, Li
name: Feng, Yingying
name: Zhou, Ying
name: Tong, Zhongkai
name: Dong, Zhaoxing
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Molecular Biology
subject: Physiology
subject: FOS: Biological sciences
subject: Pharmacology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences

description: Additional file 1: Figure S1 BLM induced PF in mice, and abnormal IL-27 expression was observed. A: HE and Masson staining to detect histological changes in the lung after 7, 14 and 28 days of BLM induction (magnification: × 200; scale bars: 100 μm); B: ELISA to detect hydroxyproline content; C, D and E: COL I and III expression in tissues by RT‒qPCR and Western blot; F and G: Detecting IL-27 expression in tissues by RT‒qPCR and Western blot. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001; values are expressed as the mean ± standard deviation (n = 5). Figure S2 IL-27 attenuates BLM-induced PF in mice. A and B: HE and Masson staining to detect histological changes in the lungs of different treatment groups after induction for 7, 14 and 28 days (magnification: × 200; scale bars: 100 μm); C, D and E: Detection of the expression of COL I and III in different treatment groups by RT‒qPCR and Western blotting. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001; values are expressed as the mean ± standard deviation (n = 5). Figure S3 Effects of IL-27 on DNMT1, ERK/p38 signaling pathways and autophagy. A: The mRNA levels of DNMT1 were detected by RT‒qPCR; B: Western blot for ERK and p38 phosphorylated protein levels; C: Western blot for LC3 and Beclin1 protein levels. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001; values are expressed as the mean ± standard deviation (n = 3). Figure S4. EdU for cell proliferation viability and immunofluorescence to detect α-SMA, FN, and COL I fluorescence intensity. A: EdU for cell proliferation viability (magnification: × 200; scale bars: 100 μm); B: Immunofluorescence to detect α-SMA, FN, and COL I fluorescence intensity (magnification: × 200; scale bars: 100 μm). Figure S5. EdU for cell proliferation viability and immunofluorescence to detect LC3, Beclin1, α-SMA, FN, and COL I fluorescence intensity. A: EdU for cell proliferation viability (magnification: × 200; scale bars: 100 μm); B: Immunofluorescence to detect LC3 and Beclin1 fluorescence intensity (magnification: × 200; scale bars: 10 μm); C: Immunofluorescence to detect α-SMA, FN, and COL I fluorescence intensity (magnification: × 200; scale bars: 100 μm). Figure S6 EdU for cell proliferation viability and immunofluorescence to detect LC3, Beclin1, α-SMA, FN, and COL I fluorescence intensity. A: EdU for cell proliferation viability (magnification: × 200; scale bars: 100 μm); B: Immunofluorescence to detect LC3 and Beclin1 fluorescence intensity (magnification: × 200; scale bars: 10 μm); C: Immunofluorescence to detect α-SMA, FN, and COL I fluorescence intensity (magnification: × 200; scale bars: 100 μm). Figure S7 EdU for cell proliferation viability and immunofluorescence to detect LC3, Beclin1, α-SMA, FN, and COL I fluorescence intensity. A: EdU for cell proliferation viability (magnification: × 200; scale bars: 100 μm); B: Immunofluorescence to detect LC3 and Beclin1 fluorescence intensity (magnification: × 200; scale bars: 10 μm); C: Immunofluorescence to detect α-SMA, FN, and COL I fluorescence intensity (magnification: × 200; scale bars: 100 μm). Figure S8 Detecting DNMT1 expression after sh-DNMT1 1–3 treatment by RT‒qPCR and Western blotting. A: Detecting DNMT1 expression after sh-DNMT1 1–3 treatment by RT‒qPCR; B: Detecting DNMT1 expression after sh-DNMT1 1–3 treatment by Western blotting. Figure S9 EdU for cell proliferation viability and immunofluorescence to detect LC3, Beclin1, α-SMA, FN, and COL I fluorescence intensity. A: EdU for cell proliferation viability (magnification: × 200; scale bars: 100 μm); B: Immunofluorescence to detect LC3 and Beclin1 fluorescence intensity (magnification: × 200; scale bars: 10 μm); C: Immunofluorescence to detect α-SMA, FN, and COL I fluorescence intensity (magnification: × 200; scale bars: 100 μm). Figure S10 Detecting LC3, Beclin1, α-SMA, FN, and COL I by RT‒qPCR and Western blotting. A: Western blot for LC3 and Beclin1 protein levels; B, C and D: RT‒qPCR detection of α-SMA, FN, and COL I mRNA expression levels; E: Western blot detection of α-SMA, FN, and COL I protein expression levels. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001; values are expressed as the mean ± standard deviation (n = 3). Figure S11 Detecting LC3, Beclin1, α-SMA, FN, and COL I by RT‒qPCR and Western blotting. A: Western blot for LC3 and Beclin1 protein levels; B, C and D: RT‒qPCR detection of α-SMA, FN, and COL I mRNA expression levels; E: Western blot detection of α-SMA, FN, and COL I protein expression levels. *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001; values are expressed as the mean ± standard deviation (n = 3).