Journal: Allergy, Asthma & Immunology Research

Article Title: The Role of Macrophage Migration Inhibitory Factor (MIF) in Asthmatic Airway Remodeling

doi: 10.4168/aair.2021.13.1.88

Figure Lengend Snippet: MIF, migration inhibitory factor; ASMC, airway smooth muscle cell; TEM, transmission electron microscope; OVA, ovalbumin; ISO-1, (S,R)3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester; DAPI, 4′,6-diamidino-2-phenylindole. * P < 0.05 vs. MIF 0 ng/mL; † P < 0.001 vs. MIF 0 ng/mL; ‡ P < 0.001 vs. control; § P < 0.001 vs. the OVA group.

Article Snippet: The MIF inhibitor ISO-1 was purchased from medChemExpress (Monmouth Junction, NJ, USA).

Techniques: Migration, Transmission Assay, Microscopy, Control

Journal: Allergy, Asthma & Immunology Research

Article Title: The Role of Macrophage Migration Inhibitory Factor (MIF) in Asthmatic Airway Remodeling

doi: 10.4168/aair.2021.13.1.88

Figure Lengend Snippet: MIF, migration inhibitory factor; ASMC, airway smooth muscle cell; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; NC, negative control; ISO-1, (S,R)3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester; q-PCR, quantitative polymerase chain reaction; shRNA, short hairpin RNA. * P < 0.001 vs. the control group; † P < 0.01 vs. the IL-4 group; ‡ P < 0.001 vs. the IL-4 group; § P < 0.01 vs. the IL-4+shRNA-NC group.

Article Snippet: The MIF inhibitor ISO-1 was purchased from medChemExpress (Monmouth Junction, NJ, USA).

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Negative Control, Real-time Polymerase Chain Reaction, shRNA, Control

Journal: Allergy, Asthma & Immunology Research

Article Title: The Role of Macrophage Migration Inhibitory Factor (MIF) in Asthmatic Airway Remodeling

doi: 10.4168/aair.2021.13.1.88

Figure Lengend Snippet: The promotive effects of macrophage-derived MIF on ASMC autophagy determined by immunofluorescence. (A) In the ISO-1-mediated MIF inhibition model, the images of GFP-RFP-LC3-transfected cells showed that ASMC autophagy was decreased in the IL-4+ISO-1 group compared with the IL-4 group (200 ×, scale bar, 10 μm). (B) In the MIF-knock-down model, the IL-4+shRNA-MIF group displayed decreased ASMC autophagy (GFP-RFP-LC3 reporting system) compared with the IL-4 or IL-4+shRNA-NC group (200 ×, scale bar, 10 μm).

Article Snippet: The MIF inhibitor ISO-1 was purchased from medChemExpress (Monmouth Junction, NJ, USA).

Techniques: Derivative Assay, Immunofluorescence, Inhibition, Transfection, Knockdown, shRNA

Journal: Cell Death & Disease

Article Title: Unveiling DEFB1 as a novel driver and promising therapeutic target in lung adenocarcinoma

doi: 10.1038/s41419-026-08748-4

Figure Lengend Snippet: A Infiltrating status of M1 (marked by F4/80 and INOS) and M2 (marked by F4/80 and CD206) macrophages in nude-mouse xenografts derived from A549 DEFB1-NC/KO/ROE cells (scale bar: 200 μm); B CoIP verification of the interaction between DEFB1 and MIF in A549/PC-9 cells treated with exogenously added 3× Flag-tagged DEFB1 (200 pg/mL); C AlphaFold 3 prediction of the interaction mode between DEFB1 and MIF; D Schematic diagram of an in vitro co-culture model of lung adenocarcinoma cells and THP-1 derived macrophages; E , F Flow Cytometry and qRT-PCR for evaluating the effects of DEFB1 and/or MIF inhibitor (ISO-1) on M1 polarization of THP-1 M0 cells co-cultured with A549 cells; G , H Flow Cytometry and qRT-PCR for evaluating the effects of DEFB1 and/or ISO-1 on M2 polarization of THP-1 M0 cells co-cultured with A549 cells; I , J Effects of DEFB1-KO, PPL-KO, and/or ISO-1 on the in vivo proliferative capacity in nude-mouse CDX model. Statistical analysis: ( F , H , and G ) One-way ANOVA test and Bonferroni correction.

Article Snippet: ISO-1 (#T3680) was obtained from TargetMol (Boston, Massachusetts, USA).

Techniques: Derivative Assay, In Vitro, Co-culture Assay, Flow Cytometry, Quantitative RT-PCR, Cell Culture, In Vivo, Analysis, Immunofluorescence, Co-Immunoprecipitation Assay, Real-time Polymerase Chain Reaction, Xenograft Assay

Journal: PLOS One

Article Title: EIF1 coordinates transcriptomic and splicing networks associated with cell cycle dysregulation in diabetic retinopathy

doi: 10.1371/journal.pone.0355473

Figure Lengend Snippet: a. Validation of EIF1 knockdown efficiency by RT-qPCR. Data are shown as mean ± SD (n = 3 per group). **** P-value < 0001. b. Validation of EIF1 knockdown efficiency by western blot. Data are shown as mean ± SD (n = 3 per group). c. Bar plot showing EIF1 expression levels (in FPKM) from RNA-seq data. Data are shown as mean ± SEM (n = 3 per group). ***P-value < 0.001. d. Principal component analysis (PCA) of samples after normalizing all genes expression levels. The ellipse for each group is the confidence ellipse. e. Volcano plot of DEGs between si-EIF1 and NC groups. Red indicates genes with up regulated expression. Blue indicates genes with down regulated expression. f. Hierarchical clustering heatmap showing expression levels of all DEGs. FPKM values are log2-transformed and then median-centred by each gene. g. Metascape enrichment analysis of upregulated (right) and downregulated (left) DEGs.

Article Snippet: The PVDF membranes were then blocked for 1h at room temprature and incubated overnight at 4°C with primary antibody respectively against EIF1 (anti-EIF1, 1:1000, antibody produced in rabbit, 15276-1-AP, Proteintech, China) and GAPDH (1:10000, antibody produced in mouse, 60004-1-lg, Proteintech, China), followed by an incubation with horseradish peroxidase-conjugated secondary antibody (anti-rabbit, 1:10000, SA00001-2, Proteintech, China or anti-mouse, 1:10000, AS003, ABclonal, China) for 45 min at room temprature.

Techniques: Biomarker Discovery, Knockdown, Quantitative RT-PCR, Western Blot, Expressing, RNA Sequencing, Transformation Assay

Journal: PLOS One

Article Title: EIF1 coordinates transcriptomic and splicing networks associated with cell cycle dysregulation in diabetic retinopathy

doi: 10.1371/journal.pone.0355473

Figure Lengend Snippet: a. The EIF1 regulates alternative splicing of COP1. Left panel: IGV-sashimi plot showing the regulated alternative splicing events and binding sites across mRNA. Reads distribution of RASE is plotted in the up panel and the transcripts of each gene are shown below. Right panel: The schematic diagrams depict the structures of ASEs. RNA-seq validation of ASEs are shown at the bottom of the right panel. Error bars represent mean ± SEM. RT-qPCR validation of COP1. Data are shown as mean ± SD (n = 3 per group). ** p < 0.01, *** p < 0.001. b. The EIF1 regulates alternative splicing of TFE3. Left panel: IGV-sashimi plot showing the regulated alternative splicing events and binding sites across mRNA. Reads distribution of RASE is plotted in the up panel and the transcripts of each gene are shown below. Right panel: The schematic diagrams depict the structures of ASEs. RNA-seq validation of ASEs are shown at the bottom of the right panel. Error bars represent mean ± SEM. RT-qPCR validation of TFE3. Data are shown as mean ± SD (n = 3 per group). * p < 0.05.

Article Snippet: The PVDF membranes were then blocked for 1h at room temprature and incubated overnight at 4°C with primary antibody respectively against EIF1 (anti-EIF1, 1:1000, antibody produced in rabbit, 15276-1-AP, Proteintech, China) and GAPDH (1:10000, antibody produced in mouse, 60004-1-lg, Proteintech, China), followed by an incubation with horseradish peroxidase-conjugated secondary antibody (anti-rabbit, 1:10000, SA00001-2, Proteintech, China or anti-mouse, 1:10000, AS003, ABclonal, China) for 45 min at room temprature.

Techniques: Alternative Splicing, Binding Assay, RNA Sequencing, Biomarker Discovery, Quantitative RT-PCR