Journal: bioRxiv

Article Title: Virus and Cell Specific HMGB1 Secretion and Subepithelial Infiltrate Formation in Adenovirus Keratitis

doi: 10.1101/2025.01.07.631509

Figure Lengend Snippet: (A) Cytoplasmic and nuclear extracts prepared from uninfected or HAdV-D37-infected for 2, 12, 24, and 48 hpi. were resolved on 4-20% SDS-PAGE. Western blots for HMGB1 in the nuclear (Nuc), cytoplasmic (Cyt), and supernatant (Sup) extracts with TBP (nuclear) β-actin (cytoplasmic) as loading controls in cell types THE, PHCE, HCF, A549, and HEK293 (left to right). (B) Densitometric analysis of HMGB1 band intensity of cytoplasmic and nuclear extracts in THE, PHCE, HCF, A549, and HEK293 (left to right). p values were determined by t test; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 ( n=3 ). (C) Quantification of extracellular HMGB1 measured from supernatants collected at indicated time points pi in THE and PHCE cells ( n=3 ). (D) Schematic representation (created using Biorender.com ) for HAdV-D37 induced translocation of HMGB1 from the cell nucleus to the cytoplasm and then into the extracellular space.

Article Snippet: A549 (CCL-185), a human lung carcinoma cell line, and HEK293 (CRL-1573), a human embryonic kidney cell line, were purchased from American Type Culture Collection (ATCC).

Techniques: Infection, SDS Page, Western Blot, Translocation Assay

Journal: bioRxiv

Article Title: Virus and Cell Specific HMGB1 Secretion and Subepithelial Infiltrate Formation in Adenovirus Keratitis

doi: 10.1101/2025.01.07.631509

Figure Lengend Snippet: (A) Western blot analysis for HMGB1 expression along with β-actin for load control from whole cell lysates of uninfected (M) or HAdV-D37-infected (V) THE cells for 2, 12, 24, and 48 hpi. qRT-PCR analysis of HMGB1 gene expression for mock and virus infected cells at the same times pi is shown as bar graphs below the Western blot. (B) Bar graph for qRT-PCR of the viral early gene E1A expression, a surrogate marker for viral entry, and normalized to human ACTG gene for quantification. (C) Viral late protein pIIIa expression in cytoplasmic and nuclear extracts prepared from uninfected or HAdV-D37-infected THE, PHCE, HCF, A549 and HEK293 for 2, 12, 24, and 48 hpi show successful infection of all cell types.

Article Snippet: A549 (CCL-185), a human lung carcinoma cell line, and HEK293 (CRL-1573), a human embryonic kidney cell line, were purchased from American Type Culture Collection (ATCC).

Techniques: Western Blot, Expressing, Control, Infection, Quantitative RT-PCR, Gene Expression, Virus, Marker