Journal: Investigative Ophthalmology & Visual Science

Article Title: Ly93 Inhibits Sphingomyelin Synthesis and Attenuates Inflammation and Injury in Dry Eye Conjunctival Organoids

doi: 10.1167/iovs.67.2.58

Figure Lengend Snippet: Hyperosmolarity induced DED-associated injury and inflammation of conjunctival organoids. ( A ) Representative images showing hyperosmolarity-induced organoid disintegration ( scale bar , 100 µm). ( B ) Quantification of the percentage of intact organoids. ( C ) Western blotting analysis of proliferation markers (p63 and Ki67), inflammatory mediators (MMP-9 and IL-1β), signaling molecules (NF-κB p65, STAT1, and PI3K), ferroptosis markers (TFR and GPX4), and apoptosis marker (caspase-3). ( D – F ) Immunofluorescence staining of p63 + and Ki67 + cells in organoids (scale bar , 50 µm). ( G ) Immunofluorescence staining of caspase-9, NF-κB p65, STAT1, and IL-1β in organoids ( scale bar , 50 µm). * P < 0.05; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: Immunofluorescence staining was performed to evaluate the expression of K19 (1:200, Cell Signaling Technology, Danvers, MA, USA), K13 (1:200, Cell Signaling Technology), IL-1β (1:200, Abcam, Cambridge, UK), STAT1 (1:200, Proteintech, Rosemont, IL, USA), p63 (1:250, Abcam), Ki67 (1:200, Abcam), NF-κB p65 (1:200, Abcam), MUC1 (1:200, Abcam), MUC5AC(1:250, Abcam), caspase-9 (1:200, Cell Signaling Technology), ZO-1 (1:200, Abcam), and 4-hydroxynonenal (4-HNE) (1:100, Invitrogen, Carlsbad, CA, USA) in frozen sections of the conjunctival organoids.

Techniques: Western Blot, Marker, Immunofluorescence, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ly93 Inhibits Sphingomyelin Synthesis and Attenuates Inflammation and Injury in Dry Eye Conjunctival Organoids

doi: 10.1167/iovs.67.2.58

Figure Lengend Snippet: Exogenous SM induced inflammation and injury of conjunctival organoids. ( A ) Effects of SM treatment on the viability of conjunctival cells and morphology of organoids ( scale bar , 100 µm for cells and 200 µm for organoids). ( B , C ) Western blotting analysis of inflammatory markers (phospho-STAT1, STAT1, IL-1β, and iNOS) in SM-treated organoids. ( D – F ) Relative mRNA expression of IL-1β, IL-6, and STAT1 after SM treatment. ( G ) Immunofluorescence staining of K13, K19, Ki67, MUC1, and MUC5AC in SM-treated organoids ( scale bar , 50 µm). ( H – L ) Comparative analysis of immunofluorescent results: K13 ( H ), K19 ( I ), Ki67 ( J ), MUC1 ( K ), and MUC5AC ( L ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: Immunofluorescence staining was performed to evaluate the expression of K19 (1:200, Cell Signaling Technology, Danvers, MA, USA), K13 (1:200, Cell Signaling Technology), IL-1β (1:200, Abcam, Cambridge, UK), STAT1 (1:200, Proteintech, Rosemont, IL, USA), p63 (1:250, Abcam), Ki67 (1:200, Abcam), NF-κB p65 (1:200, Abcam), MUC1 (1:200, Abcam), MUC5AC(1:250, Abcam), caspase-9 (1:200, Cell Signaling Technology), ZO-1 (1:200, Abcam), and 4-hydroxynonenal (4-HNE) (1:100, Invitrogen, Carlsbad, CA, USA) in frozen sections of the conjunctival organoids.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ly93 Inhibits Sphingomyelin Synthesis and Attenuates Inflammation and Injury in Dry Eye Conjunctival Organoids

doi: 10.1167/iovs.67.2.58

Figure Lengend Snippet: Ly93 attenuated DED-associated inflammation in conjunctival organoids. ( A ) Immunofluorescence staining of caspase-9, ZO-1, IL-1β, STAT1, and NF-κB p65 between vehicle control, DED, and DED + Ly93 groups ( scale bar , 50 µm). ( B ) Intracellular Fe 2+ analysis ( scale bar , 50 µm). ( C ) Immunofluorescence staining of 4-HNE ( scale bar , 50 µm). ( D – H ) Western blotting analysis of GPX4 ( D ), TFR ( E ), AKT pathway ( F ), caspase-9 and NF-κB p65 pathway ( G ), and IL-1β and STAT1 pathway ( H ) protein expression.

Article Snippet: Immunofluorescence staining was performed to evaluate the expression of K19 (1:200, Cell Signaling Technology, Danvers, MA, USA), K13 (1:200, Cell Signaling Technology), IL-1β (1:200, Abcam, Cambridge, UK), STAT1 (1:200, Proteintech, Rosemont, IL, USA), p63 (1:250, Abcam), Ki67 (1:200, Abcam), NF-κB p65 (1:200, Abcam), MUC1 (1:200, Abcam), MUC5AC(1:250, Abcam), caspase-9 (1:200, Cell Signaling Technology), ZO-1 (1:200, Abcam), and 4-hydroxynonenal (4-HNE) (1:100, Invitrogen, Carlsbad, CA, USA) in frozen sections of the conjunctival organoids.

Techniques: Immunofluorescence, Staining, Control, Western Blot, Expressing

Journal: bioRxiv

Article Title: Integrating Artificial Intelligence and Bioinformatics Methods to Identify Disruptive STAT1 Variants Impacting Protein Stability and Function

doi: 10.1101/2024.10.04.616446

Figure Lengend Snippet: Selection of nsSNPs of STAT1 gene.

Article Snippet: Identification of nsSNPs in STAT1 protein domains The FASTA sequence of the STAT1 protein was submitted to the InterPro server ( https://www.ebi.ac.uk/interpro ).

Techniques: Selection

Journal: Endocrine-related cancer

Article Title: Signal transducer and activator of transcription 5A/B in prostate and breast cancers

doi: 10.1677/ERC-08-0013

Figure Lengend Snippet: Chromosomal mapping and sequence identity between human and mouse signal transducer and activator of transcriptions (STATs)

Article Snippet: In the mouse, Stat1 and Stat4 are located on chromosome 1 (band 1 C1.1) ( Schindler et al . 1992 , Yamamoto et al . 1994 ); Stat2 and Stat6 are on chromosome 10 (band 10 D3) ( Fu et al . 1992 , Quelle et al . 1995 ); and Stat3 , Stat5a , and Stat5b map to chromosome 11 (band 11 D) ( Zhong et al . 1994 a , Copeland et al . 1995 , Shi et al . 1996 , Levy et al . 1998 ; ). table ft1 table-wrap mode="anchored" t5 caption a7 STAT Human Mouse % Identity References Amino acid Chromosome Amino acid Chromosome STAT1 750 2 749 1 93 Schindler et al . (1992) and Haddad et al . (1998) STAT2 846 12 915 10 64 Fu et al . (1992) and Goureau et al . (2001) STAT3 769 17 769 11 99 Zhong et al . (1994 a ) and Choi et al . (1996) STAT4 748 2 748 1 94 Yamamoto et al . (1994 , 1997) STAT5A 794 17 793 11 96 Mui et al . (1995) and Lin et al . (1996) STAT5B 787 17 786 11 96 Mui et al . (1995) and Lin et al . (1996) STAT6 847 12 837 10 85 Quelle et al . (1995) and Leek et al . (1997) Open in a separate window Chromosomal mapping and sequence identity between human and mouse signal transducer and activator of transcriptions (STATs) STATs are proteins of 750–900 amino acids (90–115 kDa) with five structurally and functionally conserved domains that allow the transduction of ligand-specific signals ( Schindler & Darnell 1995 ).

Techniques: Sequencing