Journal: bioRxiv

Article Title: Wnt/β-catenin signaling promotes zebrafish osteoblast dedifferentiation by wnt10a -mediated inhibition of NF-κB

doi: 10.64898/2025.12.29.696582

Figure Lengend Snippet: A) RNASeq (left) and RT-qPCR (right) reveal upregulation of the NF-κB signaling target genes nfkbiaa and nfkbiab in osteoblasts sorted from bglap:GFP fish heterozygous for the wnt10a mutation relative to their wild-type siblings at 1 dpa. nE (qPCR biological replicates) = 3, nA = 10 per replicate. ΔΔCt values are normalized to the mean of the wildtype at 1 dpa. Error bars, Mean ± SEM. Two tailed Student’s t-test. B) Osteoblast dedifferentiation, as measured by bglap downregulation in segment -1 revealed by HCR in situ hybridization, is inhibited in fish treated with the Wnt inhibitor IWR-1, while IP injection of the NF-κB inhibitor Bay-11 slightly but significantly enhances dedifferentiation. Treatment with both inhibitors yields results similar to those of Bay-11 alone. nE = 3 (except for 2 for IWR-1), nA = 18 total per group (12 for IWR-1), nR = 33 (DMSO), 24 (IWR-1), 37 (Bay-11), 33 (both). C) Overexpression of wnt10a using hs:wnt10a fish is sufficient to cause downregulation of bglapl detected by HCR in situ hybridization in non-injured fins, relative to heat-shocked wild-type fish. HCR signals were quantified in a bony segment (“B”) that is located at the same proximal-distal position as “segment -1” in amputated fins. nE = 2, nA = 12 total per group, nR = 22 total per group. Dashed line, joints. Scale bar, 100 µm. D) Immunofluorescence on cryosections of hs:wnt10a transgenic hearts reveals increased embryonic myosin heavy chain (embMHC) expression in Myl7+ cardiomyocytes at the wound border at 7 days post injury (dpi). Plots show the ventricular area covered by anti-embMHC staining relative to the 150 µm wound border zone area occupied by Myl7+ myocardium. nE = 2, nA = 13 wild-type, 11 hs:wnt10a . Scale bar, 100 µm. (B, C, D) Error bars, mean ± 95% CI. Two tailed Student’s t-test.

Article Snippet: Primary antibodies against embMHC Mouse monoclonal (MYH7 DSHB, N2.261, RRID:AB_531790), and Myl7 Rabbit polyclonal (GeneTex, GTX128346, RRID:AB_2885759) were diluted in PEMTx/normal goad serum and applied overnight at 4 °C.

Techniques: Quantitative RT-PCR, Mutagenesis, Two Tailed Test, In Situ Hybridization, Injection, Over Expression, Immunofluorescence, Transgenic Assay, Expressing, Staining

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A-B. Group mean +/- SEM percent of fluorescent radiant efficiency (p/s)/(µW/cm 2 ) relative to radiant efficiency at 24 hr timepoint measured at site of injection for 50-, 100- or 200-cycle ALD coated powders administered at 2 mg/mL or (B) 0.4 mg/mL. Unconjugated, liquid IVISense680 fluorescent dye, conjugated OVA-IVISense680 or Alhydrogel-adsorbed, conjugated OVA-IVISense680 were administered at equimolar concentrations to amount of fluorescent dye in ALD coated powder at 2 mg/mL dose. Non-linear 4PL regression fit line with 95% confidence intervals shown for ALD coated powders (constraints on 4PL regression bottom = 0, top = 100) C. Correlation of time to 50% particle dissolution in vitro and time to 50% fluorescent signal loss in vivo (using 4PL fit parameters) at indicated doses. Simple linear regression of data at each dose shown, with slope of regression line indicated on plot. D-E. Anti-OVA IgG1 seroconversion percentage following vaccination with 100-cycle ALTA ® OVA dosed at 2 mg/mL (960 ng OVA) or 0.4 mg/mL (192 ng OVA) or (E) 200-cycle ALTA ® OVA dosed at 2 mg/mL (920 ng OVA) or 0.4 mg/mL (192 ng OVA). The IgG1 data is overlaid with group mean +/- SEM percent of fluorescent radiant efficiency relative to 24 hr timepoint measured at site of injection. Dotted lines at week 2 or week 7 indicate shift in rate of fluorescent signal loss for (D) 100-cycle or (E) 200-cycle powders, respectively.

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection, Dissolution, In Vitro, In Vivo

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A. Study design, n=10 mice per group. All mice received 62.5 ng OVA dose, either in a single injection, or over the course of 7 daily injections. Of note, Al 3+ ion content differs significantly between groups treated with ALD-coated material or Alhydrogel. B. Anti-OVA IgG1 titers at week 3 post first injection (14 days post final injection for 7 day dose groups). Plot shows geometric mean titer (n=10/group) with 95% confidence interval, **** = p < 0.0001. Data analyzed using Kruskal-Wallis test. C. Anti-OVA IgG1 titers throughout study, plot shows geometric mean titer (n=10/group) with 95% confidence interval. D. Anti-OVA IgG1 seroconversion percentage, indicating a 2 log-fold increase in anti-OVA IgG1 titers relative to pre-injection baseline.

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A. Anti-OVA IgG1 titers following administration of 200 ng OVA dose given from Alhydrogel-adsorbed OVA liquid prime/boost (injection schedule 100 ng D0/100 ng D28 or 100 ng D0/100 ng D49). For mixed products (green/purple traces) 100- or 200-cycle ALTA ® powders containing a 100 ng OVA dose were resuspended in a diluent containing 100 ng Alhydrogel-adsorbed OVA and given as a single injection on D0. Plots shows geometric mean titer (n=8-10/group) with 95% confidence interval. B. Total AUC of log10-transformed anti-OVA IgG1 titers shown in A from week 0 to week 16. Column shows mean AUC +/- SEM (n=8-10 mice/group) Data analyzed using one-way ANOVA with Tukey’s multiple comparisons test. **** = p < 0.0001. *** = p < 0.001. ** = p < 0.01. C. The percentage of OVA-specific CD8+ T cells relative to total CD8+ T cells in whole blood was measured using flow cytometry (see methods). Plot shows mean +/- SEM (n=5 mice/group). D. Anti-OVA IgG1 titers following a 200 ng OVA dose given in 50-, 100- or 200-cycle ALD coated vaccine powder. For the mixed ALD vaccine product (purple), 50-cycle and 200-cycle materials were resuspended in diluent at a 2x concentration, then mixed at a 1:1 ratio immediately prior to injection to deliver a total dose of 200 ng OVA. Plot shows geometric mean titer (n=9-10/group) with 95% confidence interval. E. Total area under the curve of anti-OVA IgG1 titer was calculated for all animals after vaccination with 200 ng OVA from 50-, 100-, 200- or mixed 50+200-cycle ALTA ® OVA powders. Plot shows the group mean AUC +/- SEM (n=9-10/group). Data analyzed using one-way ANOVA with Tukey’s multiple comparisons test. * = p < 0.05. F. The percentage of OVA-specific CD8+ T cells relative to total CD8+ T cells in whole blood was measured using flow cytometry (see methods). Plot shows mean +/- SEM (n=5 mice/group).

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection, Transformation Assay, Flow Cytometry, Concentration Assay

Journal: bioRxiv

Article Title: Atomic Layering Thermostable Antigen and Adjuvant (ALTA ® ) platform provides unique antigen delivery system through controlled release to improve immune response to vaccination

doi: 10.64898/2026.01.05.697591

Figure Lengend Snippet: A. Total anti-N332-GT5 gp140 IgG titers after a single administration of 50-cycle ALTA ® on D0 at indicated doses of N332-GT5 gp140. Plots shows geometric mean titer (n=8/group) +/- 95% confidence interval. B. Analysis of fluorescent signal at site of injection following administration of 50-cycle ALTA ® containing fluorescently-labeled N332-GT5 gp140. Plot shows mean +/- SEM of percent of fluorescent radiant efficiency (p/s)/(µW/cm 2 ) relative to radiant efficiency at 24 hr timepoint measured at site of injection for 50-cycle ALTA ® N332-GT5 gp140 administered at specified doses (square traces). Total anti-N332-GT5 gp140 IgG titers plotted as geometric mean +/- 95% confidence interval (n=5 mice/group) (triangle traces). C. Kinetics of total anti-N332-GT5 gp140 IgG titers elicited by a single administration of 10 µg N332-GT5, delivered in a liquid formulation (blue) or in 50-cycle ALTA ® products with/without the presence of adjuvants in the injection diluent. Plot shows geometric mean titer +/- 95% confidence interval (n=16 mice/group red trace, n=8 mice/group all others) D. Total anti-N332-GT5 gp140 IgG1, IgG2b, IgG2c and IgG3 titers elicited by a single administration of 10 µg N332-GT5 gp140 at week 8 post injection, delivered in a liquid formulation (blue) or in 50-cycle ALTA ® products with/without the presence of adjuvants in the injection diluent. Plot shows geometric mean titer +/- 95% confidence interval (n=16 mice/group red trace, n=8 mice/group all others). ** = p < 0.01, * = p < 0.05, data analyzed using Kruskal-Wallis test. E-H. Total anti-N332-GT5 gp140 (E) IgG1, (F) IgG2b, (G) IgG2c and (H) IgG3 titers elicited by a single administration of 10 µg N332-GT5 gp140 at week 8 post injection, delivered in a liquid formulation (blue) or in 50-cycle ALTA ® products with/without the presence of adjuvants in the injection diluent. Plot shows geometric mean titer +/- 95% confidence interval (n=16 mice/group red trace, n=8 mice/group all others. **** = p < 0.0001, ** = p < 0.01, * = p < 0.05, ns = p > 0.05, data analyzed using Kruskal-Wallis test.

Article Snippet: A standard curve was established with a dilution series of mouse anti-OVA IgG1 primary antibody (Chondrex #7093).

Techniques: Injection, Labeling, Formulation

Journal: BMC Pulmonary Medicine

Article Title: Optimization of intranasal bleomycin dose for effective pulmonary fibrosis induction in mice with minimal animal distress

doi: 10.1186/s12890-025-04001-4

Figure Lengend Snippet: Histopathological characterization in BLM-treated animals (IN—5 mg/kg). A ) Histopathological evaluation by H&E staining (upper panel), Sirius Red staining (middle panel), and Masson’s trichrome (lower panel) of lungs of vehicle mice (CTR) or treated with BLM after a single IN administration. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 500 µm (upper panel) – 100 µm (lower panels). B ) IHC for Iba1 (left panel), CD68 (green—middle panel), and α-SMA staining (right panel) of lungs of CTR or treated with BLM. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported, scale bar = 100 µm. The boxed areas (Iba1 black, CD68 red, α-SMA green) in CTR and day 7 sections are shown at higher magnification in the right panel, scale bar = 50 µm. C) Histopathological quantification of Sirius Red staining in lung section (left graph) and representation of Ashcroft scale grade (right graph) obtained by Masson’s trichrome analysis of lungs of CTR or treated with BLM. Data are reported as mean ± SE. The data were analyzed by Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, ** p ≤ 0.01. D ) mRNA expression of TNF-α, COL 1a1, and FN1 was evaluated by RT-qPCR in the lungs of mice ( n = 3 per group) treated with BLM and sacrificed at different time points. Genes were normalized on β-ACT, and the 2 −ΔΔCt method was employed for relative quantification on an external calibrator. Data are reported as mean ± SE and were analyzed with Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01. E ) SMAD 2/3 and pSMAD 2 expression in the lungs from CTR mouse at days 7, 14, 21, and 28 of treatment obtained with WB

Article Snippet: For subcellular localization, primary rat anti-CD68 monoclonal antibody (1:200, Serotec, Kidlington, UK) + Triton X-100 0.1% + NGS 3% in 1X PBS overnight at 4 °C was used.

Techniques: Staining, Expressing, Quantitative RT-PCR, Quantitative Proteomics

Journal: BMC Pulmonary Medicine

Article Title: Optimization of intranasal bleomycin dose for effective pulmonary fibrosis induction in mice with minimal animal distress

doi: 10.1186/s12890-025-04001-4

Figure Lengend Snippet: Histopathological characterization in BLM-treated animals (IN—3 mg/kg). A Histopathological evaluation by H&E staining (upper panel), Sirius Red staining (middle panel), and Masson’s trichrome (lower panel) of lungs of vehicle mice (CTR) or treated with BLM after a single IN administration. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 500 µm (upper panel) – 100 µm (lower panels). B IHC for Iba1 (left panel), CD68 (green—middle panel), and α-SMA staining (right panel) of lungs of CTR or treated with BLM. Representative images of lung sections of animals sacrificed at 7, 14, 21, and 28 days after the treatment are reported. Scale bar = 100 µm. The boxed areas (Iba1 black, CD68 red, α-SMA green) in CTR and day 7 sections are shown at higher magnification in the right panel, scale bar = 50 µm C ) Histopathological quantification of Sirius Red staining in lung section (left graph) and representation of Ashcroft scale grade (right graph) obtained by Masson’s trichrome analysis of lungs of CTR or treated with BLM. Data are reported as mean ± SE. The data were analyzed by Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. D ) mRNA expression of TNF-α, COL 1a1, and FN1 was evaluated by RT-qPCR in the lungs of mice ( n = 3 per group) treated with BLM and sacrificed at different time points. Genes were normalized on β-actin, and the 2 −ΔΔCt method was employed for relative quantification on an external calibrator. Data are reported as mean ± SE and were analyzed with Kruskal–Wallis test followed by Dunn’s test. Significant differences compared to the CTR are reported, * p ≤ 0.05, ** p ≤ 0.01. E) SMAD 2/3 and pSMAD 2 expression in the lungs from CTR mouse at days 7, 14, 21, and 28 of treatment obtained with WB

Article Snippet: For subcellular localization, primary rat anti-CD68 monoclonal antibody (1:200, Serotec, Kidlington, UK) + Triton X-100 0.1% + NGS 3% in 1X PBS overnight at 4 °C was used.

Techniques: Staining, Expressing, Quantitative RT-PCR, Quantitative Proteomics