Journal: The EMBO Journal

Article Title: Arthropod exosomal glycine-rich protein as a potential vaccine candidate effectively reduces tick blood-feeding and pathogen transmission

doi: 10.1038/s44318-026-00709-z

Figure Lengend Snippet: Hematoxylin and eosin (H&E) staining of skin biopsy samples from mice (that allowed feeding of mock/ XM_002400035 -dsRNA-treated ticks) display that tick feeding causes inflammation at bite site in mock-dsRNA-treated group ( A ), but inflammation is reduced in XM_002400035 -dsRNA-treated group ( B ). Within the panniculus, there is downward projection of epidermis containing chitinous tick mouthparts (shown by black arrow). The panniculus contained moderate to large number of neutrophils, lymphocytes, plasma cells, and lower number of macrophages in mice that allowed feeding of mock-dsRNA-treated ticks ( A ). However, panniculus contained moderate number of inflammatory cells (shown by black arrow) with lymphocytes mixed with few macrophages and plasma cells in mice that allowed feeding of XM_002400035 -dsRNA-treated ticks. There is a mild crush artifact in this image. Magnification of both these images is 200×. Scale bar indicates 100 μm for each image. Enlarged images shown in Fig. 9A,B are repeated in Appendix Fig. for better visualization. ( C ) ELISA assay performed with skin lysates from mice that allowed feeding of ticks silenced for exosomal GRP or mock control ticks. Samples were probed with serum from immunized mice (1:1000 dilution). ( D ) Scratch assays performed on HaCaT cell monolayers incubated with 2 µg of GST/GST- GRP/GST-CXCL-12 protein (for 12 h), with/without 20 µl of tick exosomes from uninfected (UI), LGTV-infected (I), LGTV-infected and mock-dsRNA-treated or LGTV-infected and XM_002400035 -dsRNA-treated groups are shown. Phase contrast images (obtained using EVOS auto-fluorescence system, M7000) of HaCaT cell monolayers were taken for selected time-points (as before scratch, 0, 16, 20, and 24 h) and using 10× magnification. Untreated (UT) monolayers served as internal control. Scale bar indicates 275 μm for each image per group or timepoint. ( E ) Measurement of remaining wound size diameters (analyzed by ImageJ software) at different time-points (of 0, 16, 20, and 24 h) post-treatment of tick exosomes-derived from UI, I, mock/ XM_002400035 -dsRNA is shown. Wounds at 0 h were considered as 100% for all groups, including untreated (UT) control. Mouse CXCL-12 expression was analyzed in skin samples from mice immunized with GST/GST-GRP protein is shown ( F ). Exact number of sample numbers for each group representing multiple experiments is 5 mice for GST/6 mice for GST-GRP groups (in C , F ). Statistical differences were calculated using Mann–Whitney U test and p value is shown. p < 0.05 is considered as statistically significant. ( G ) Schematic model showing tick-borne flavivirus transmission to vertebrate host via tick saliva-derived exosomes. Ixodes scapularis tick attaches firmly and bites on host skin for longer feeding. Secreted saliva contains a plethora of substances including cement and perhaps cement-like GRPs to seal the feeding cone/cavity for directional blood flow and to defend from being groomed off by the vertebrate host. During blood meal ingestion, infected-ticks may continuously spit saliva containing infectious exosomes with viral full-length RNA genomes or polyproteins at host skin interface. We propose that incubation of tick exosomes containing exosomal GRP modulates the battle ground at skin interface by delaying cell migration/recruitment of immune cells (like neutrophils and dendritic cells from circulation) at the wound/bite site. Tick exosomes containing GRP inhibits residential keratinocytes and IL-8/CXCL-12 to delay injury, wound-healing, tissue damage, and repair process that will eventually enable ticks to acquire a successful blood meal at the host skin interface. .

Article Snippet: Keratinocyte growth kit , ATCC , # PCS-200-040.

Techniques: Staining, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Incubation, Infection, Fluorescence, Software, Derivative Assay, Expressing, MANN-WHITNEY, Transmission Assay, Migration

Journal: Toxicology and applied pharmacology

Article Title: Monoubiquitinated γ-H2AX: abundant product and specific biomarker for non-apoptotic DNA double-strand breaks

doi: 10.1016/j.taap.2018.07.007

Figure Lengend Snippet: Panels A-C used antibodies #1. (A) Impact of gel polyacrylamide percentage and transfer conditions to PVDF membranes. Treatments of H460 cells: 6 h with 3 mM hydroxyurea (HU), 6 or 16 h with 5 μM aphidicolin (Aphi) or 1 h with 5 μM bleomycin (Bleo). All γ-H2AX images had 5 s film exposures. (B) Ethanol effect in semi-dry transfer. IMR90 cells were treated with 40 μM etoposide (Eto) for 1 h. (C) Comparison of wet (pH 8.2 and pH 8.8) and semi-dry transfers using 12% ethanol-containing buffers (10 s film exposures for all γ-H2AX images). IMR90 cells were treated with 40 μM etoposide for 1 h. (D) Effectiveness of different antibodies using semi-dry transfer with 12% ethanol. Normal human keratinocytes were treated for 1 h with 30 μM etoposide or 0.5 μM bleomycin. Antibodies #1–3: 1:1000 dilutions and 5 s exposures, antibody #4: 1:1000 dilution and 60 s exposure. (E) Comparison of antibodies #1 and #5 (both at 1:1000 dilutions) in H460 cells treated with bleomycin for 1 h. (F) γ-H2AX blot (Ab#1) in control and H2AX-depleted IMR90 cells treated with 100 μM etoposide for 1h.

Article Snippet: IMR90, WI38, normal human neonatal keratinocytes (PCS-200–010), H460, A549, U2OS, mouse embryonic fibroblasts (MEFs, SCRC-1040), telomerase-immortalized human bronchial epithelial HBEC3 (CRL-4051), Daudi, HCT116, Ramos, Raji and CA46 were obtained from ATCC.

Techniques: Comparison, Control

Journal: bioRxiv

Article Title: KGF induces podosome formation via integrin-Erk1/2 signaling in human immortalized oral epithelial cells

doi: 10.1101/508416

Figure Lengend Snippet: KGF Mediated its Effects on HIOECs through Erk1/2 and Akt Signaling. (A and B) Western blot detection of the expression level of KGFR in HIOECs after KGF treatment, (C and D) the phosphorylation level of Jak2 and Stat3 after KGF stimulation for 15 min, and (E and F) the phosphorylation level of Erk1/2, p38, Jnk, and Akt after KGF stimulation for 15 min. n=3 independent experiments. Analysis of matrix protein expression in protein extracts from each group by ImageJ. (G) EdU labeling assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. (H) The quantification shown in the right graph indicates the mean ± SEM. (I) MTT assay: the impact of U0126 and Ly294002 on the proliferation ability of HIOECs. Data are expressed as the means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: HIOECs at 3x10 4 cells/well were seeded in the 96-well plate and exposed to KGF (10 ng/mL; Proteintech) for 24 h. For MTT assays, the cells were incubated with 20 μL of MTT solution (5 mg/mL) at 37°C for 4 h. The supernatant was discarded, and 150 μL of dimethyl sulfoxide (DMSO) was added to each well.

Techniques: Western Blot, Expressing, Phospho-proteomics, Labeling, MTT Assay