Journal: Biochemistry and Biophysics Reports

Article Title: Dual knockdown of Alox15 and TGF-β1 by lipid nanoparticle-delivered siRNA in bleomycin-induced pulmonary fibrosis

doi: 10.1016/j.bbrep.2026.102534

Figure Lengend Snippet: RNA sequencing analysis of an idiopathic pulmonary fibrosis dataset revealed upregulation of ALOX15 and TGF-β1 mRNA compared to controls. RNA sequencing was performed to compare the expression of ALOX15 and TGF-β1 between idiopathic pulmonary fibrosis and normal samples from GSE134692 and GSE150910 datasets.

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against Alox15 (ab244205, Abcam), 4-HNE (ab48506, Abcam), TGF-β1 (sc-130348, Santa Cruz Biotechnology, USA), E-cadherin (IR55-180, iReal Biotechnology), Vimentin (GTX100619, GeneTex, Taiwan), α-SMA (E-AB-34268, Elabscience, China), or GAPDH (2118, Cell Signaling Technology, Danvers, MA, USA).

Techniques: RNA Sequencing, Expressing

Journal: Biochemistry and Biophysics Reports

Article Title: Dual knockdown of Alox15 and TGF-β1 by lipid nanoparticle-delivered siRNA in bleomycin-induced pulmonary fibrosis

doi: 10.1016/j.bbrep.2026.102534

Figure Lengend Snippet: Bleomycin-induced IPF is associated with elevated lung Alox15 and TGF-β1 expression. Mice were administered with bleomycin (BLM) or saline on day 0. The mice in the BLM group were sacrificed on days 2, 4, 7 or 14 after treatment, and lung specimens were harvested. n = 6 per group. (A) Lung function tests were performed on day 14 before sacrifice. Rrs, airway resistance; Ers, lung tissue elastance; Cst, Static lung compliance; IC, Inspiratory capacity. (B) Histological analysis with H&E staining and Picro Sirius Red staining. Scale bar = 200 μm. (C) Quantification of Picro Sirius Red staining. Scatter plot showing the percentage of the positive area of staining in lung tissue. (D) Quantification of hydroxyproline concentration. (E) Real-time PCR were performed to detect the expression of Alox15 and TGF-β1 in the pulmonary tissues from each group. (F) Western blot analysis and (G) IHC analysis and quantification to detect Alox15, 4-HNE, and TGF-β1 expression. Scale bar = 50 μm. Scatter plot showing the percentage of the positive area in lung tissue. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against Alox15 (ab244205, Abcam), 4-HNE (ab48506, Abcam), TGF-β1 (sc-130348, Santa Cruz Biotechnology, USA), E-cadherin (IR55-180, iReal Biotechnology), Vimentin (GTX100619, GeneTex, Taiwan), α-SMA (E-AB-34268, Elabscience, China), or GAPDH (2118, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Saline, Staining, Concentration Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Dual knockdown of Alox15 and TGF-β1 by lipid nanoparticle-delivered siRNA in bleomycin-induced pulmonary fibrosis

doi: 10.1016/j.bbrep.2026.102534

Figure Lengend Snippet: LNP-mediated delivery of siRNAs into lung epithelial and fibroblast cells effectively reduces mRNA levels both in vitro and in vivo . (A, B) MLE-12 cells and murine lung fibroblasts were treated with negative control (NC), Alox15, or TGF-β1 siRNA for 24 h, followed by treatment with bleomycin for an additional 24 h, then subjected to real-time PCR assay. Fluorescence images of (C) MLE-12 cells and (D) murine lung fibroblasts treated with 50 nM naked siRNA-Cy5 or siRNA-Cy5 encapsulated with LNPs (siRNA@LNP) for 6 h, followed by staining with Hoechst 33342. Scale bar = 20 μm. (E) Flow cytometry analysis showing the mean fluorescent intensity (MFI) of Cy5 positive cells. (F) Mice were injected with saline, naked siRNA, or siRNA@LNP and sacrificed at 24 h after treatment. Cy5 fluorescence signal in the heart, lung, liver, spleen and kidney was measured by The PhotonIMAGER Optima system immediately. Fluorescence intensity in the lung expressed as total radiant efficiency. n = 3 per group. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Scatter plots representing individual mice.

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against Alox15 (ab244205, Abcam), 4-HNE (ab48506, Abcam), TGF-β1 (sc-130348, Santa Cruz Biotechnology, USA), E-cadherin (IR55-180, iReal Biotechnology), Vimentin (GTX100619, GeneTex, Taiwan), α-SMA (E-AB-34268, Elabscience, China), or GAPDH (2118, Cell Signaling Technology, Danvers, MA, USA).

Techniques: In Vitro, In Vivo, Negative Control, Real-time Polymerase Chain Reaction, Fluorescence, Staining, Flow Cytometry, Injection, Saline

Journal: Biochemistry and Biophysics Reports

Article Title: Dual knockdown of Alox15 and TGF-β1 by lipid nanoparticle-delivered siRNA in bleomycin-induced pulmonary fibrosis

doi: 10.1016/j.bbrep.2026.102534

Figure Lengend Snippet: Administration of Alox15 and TGF-β1 siRNA encapsulated with LNPs prevented pulmonary fibrosis and improved lung function in bleomycin-induced IPF mice. Mice were administered bleomycin (BLM) or saline on day 0 and treated with siRNA encapsulated LNPs (si-NC@LNP, si-Alox15@LNP, si-TGF-β1@LNP, or si-Alox15/si-TGF-β1@LNP) on days 1, 4, and 7. The mice were sacrificed on day 14, and lung specimens were harvested. n = 6 per group. (A) Lung function tests were performed before sacrifice. Rrs, airway resistance; Ers, lung tissue elastance; Cst, Static lung compliance; IC, Inspiratory capacity. (B) Histological analysis with H&E staining and Picro Sirius Red staining. Scale bar = 200 μm. (C) Quantification of Picro Sirius Red staining. Scatter plot showing the percentage of the positive area of staining in lung tissue. (D) Quantification of hydroxyproline concentration. (E) Real-time PCR was performed to detect the expression of Alox15 and TGF-β1 in the pulmonary tissues from each group. (F) Western blot analysis and (G) IHC analysis and quantification to detect Alox15, 4-HNE, and TGF-β1 expression. Scale bar = 50 μm. Scatter plot showing the percentage of the positive area in lung tissue. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against Alox15 (ab244205, Abcam), 4-HNE (ab48506, Abcam), TGF-β1 (sc-130348, Santa Cruz Biotechnology, USA), E-cadherin (IR55-180, iReal Biotechnology), Vimentin (GTX100619, GeneTex, Taiwan), α-SMA (E-AB-34268, Elabscience, China), or GAPDH (2118, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Saline, Staining, Concentration Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot