Journal: Laboratory Animal Research

Article Title: Fludarabine attenuates inflammation and dysregulated autophagy in alveolar macrophages via inhibition of STAT1/IRF1 pathway

doi: 10.1186/s42826-025-00245-7

Figure Lengend Snippet: The STAT1/IRF1 pathway is associated with LPS-induced ALI in mice. A RNA-seq data presenting comparisons of the expression of several genes, including IRF1 family genes, between the control and LPS groups of mouse lung tissue lysates ( n = 2 per group). B Expression of IRF family genes was measured as FPKM values. C Western blotting of IRF1 and IRF7 in mouse lung tissue (upper) and their densitometric ratios against β-actin (bottom) ( n = 3 per group). * p < 0.05 compared with the control group. ns: not significant. D Confocal images of immunostaining for IRF1 (green) in BAL cells. Scale bar, 10 μm ( n = 2 per group). E Western blotting of pSTAT1 (Tyr701), pSTAT1 (Ser727), total STAT1, and β-actin in mouse lung tissue (upper) and their densitometric ratios against STAT1 (bottom) ( n = 3 per group). The data are presented as mean ± SD

Article Snippet: Primary antibodies against pSTAT1 (Ser727), pSTAT1 (Tyr701), STAT1, IRF1, IRF7, LC3B, and β-actin were purchased from Cell Signaling Technology, (Danvers, MA, USA).

Techniques: RNA Sequencing, Expressing, Control, Western Blot, Immunostaining

Journal: Laboratory Animal Research

Article Title: Fludarabine attenuates inflammation and dysregulated autophagy in alveolar macrophages via inhibition of STAT1/IRF1 pathway

doi: 10.1186/s42826-025-00245-7

Figure Lengend Snippet: The STAT1/IRF1 inhibitor Fluda attenuates LPS-induced ALI in mice. A pSTAT1 (Tyr701), pSTAT1 (Ser727), STAT1, IRF1, and β-actin were detected in mouse lung tissue. B Densitometric ratios of pSTAT1 (Tyr701) and pSTAT1 (Ser727) against STAT1 and that of IRF1 against β-actin. C pSTAT1 (Tyr701), pSTAT1 (Ser727), STAT1, IRF1, and β-actin expression was detected in BAL cell lysates using western blotting. D Densitometric ratios of pSTAT1 (Tyr701) and pSTAT1 (Ser727) against STAT1 and that of IRF1 against β-actin ( n = 3). E Representative images of hematoxylin and eosin-stained lung sections from four experimental groups (× 200). Scale bar, 50 μm. F BAL cells were subjected to Giemsa staining and then observed under a microscope (× 200). Scale bar, 50 μm. G The lung injury score illustrated Fluda reduced LPS-induced ALI in mice ( n = 4 per group). H The numbers of macrophages and neutrophils in BALF. I Total protein in BALF was measured using the BCA assay. J MPO activity was measured in whole-lung lysates. K Inflammatory cytokines in BALF, including TNF-α, IL-6, IFN-γ, and IL-1β, were detected using ELISA. The data are presented as mean ± SD, n = 3 independent experiments were performed. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the control group. † p < 0.05, †† p < 0.01, and ††† p < 0.001 compared with the LPS group

Article Snippet: Primary antibodies against pSTAT1 (Ser727), pSTAT1 (Tyr701), STAT1, IRF1, IRF7, LC3B, and β-actin were purchased from Cell Signaling Technology, (Danvers, MA, USA).

Techniques: Expressing, Western Blot, Staining, Microscopy, BIA-KA, Activity Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Laboratory Animal Research

Article Title: Fludarabine attenuates inflammation and dysregulated autophagy in alveolar macrophages via inhibition of STAT1/IRF1 pathway

doi: 10.1186/s42826-025-00245-7

Figure Lengend Snippet: STAT1/IRF1, iNOS, and NF-κB/ERK1/2 activation was attenuated by Fluda in RAW264.7 cells. A pSTAT1 (Tyr701), pSTAT1 (Ser727), IRF1, and STAT1 were detected in RAW264.7 cells. STAT1 and β-actin were used as loading controls. Densitometric ratios of pSTAT1 (Tyr701) and pSTAT1 (Ser727) against STAT1 and that of IRF1 against β-actin. B Relative Nos2 expression in RAW264.7 cells was measured using real-time PCR. C Western blotting for iNOS expression. Densitometric ratio of iNOS against β-actin. D NO release was determined by measuring the amount of nitrite in conditioned medium using Griess reagent. E IL-6 and ( F ) TNF-α levels in conditioned medium were detected using ELISA. G Western blotting for p-NF-κB (Ser536), p-ERK1/2, p-JNK1/2, and p-p38. Total NF-κB, total JNK1/2, total ERK1/2, total p38, and β-actin were used as loading controls. All blots were subjected to densitometric analysis and relative quantification. Data are presented as mean ± SD ( n = 3 per group). * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the control group. † p < 0.05, †† p < 0.01, and ††† p < 0.001 compared with the LPS group. ns: not significant

Article Snippet: Primary antibodies against pSTAT1 (Ser727), pSTAT1 (Tyr701), STAT1, IRF1, IRF7, LC3B, and β-actin were purchased from Cell Signaling Technology, (Danvers, MA, USA).

Techniques: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative Proteomics, Control