Journal: iScience

Article Title: Cardiomyocyte-derived HSPB1 regulates TGF-β1 maturation and inhibits endothelial-to-mesenchymal transition in myocardial fibrosis

doi: 10.1016/j.isci.2026.115028

Figure Lengend Snippet: Regulatory role of HSPB1 in endothelial cell EndoMT (A) Western blot shows HSPB1 expression in HUVECs following lentiviral-mediated overexpression (LV-HSPB1) or knockdown (LV-HSPB1-RNAi); β-actin served as a loading control. (B) Quantification of HSPB1/β-actin ratio shows significant differences between groups. (C) Representative images of Transwell migration assays evaluating the effect of HSPB1 on TGF-β1–induced endothelial migration (scale bars, 100 μm). (D) Quantification of migrated cells per field. (E) Representative tube formation images showing the effect of HSPB1 modulation on TGF-β1–induced angiogenic activity (scale bars, 200 μm). (F–H) Quantitative analysis of tube formation parameters, including the number of branches (F), loops (G), and total tube length (H), measured using ImageJ software. Data are presented as mean ± SD ( n ≥ 6). Exact p values are indicated in the graphs. Statistical analyses were performed using one-way ANOVA followed by a Bonferroni post hoc test.

Article Snippet: Human umbilical vein endothelial cells (HUVECs; primary, pooled donors; ATCC) were purchased through an authorized distributor in China and originally sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Western Blot, Expressing, Over Expression, Knockdown, Control, Migration, Activity Assay, Software

Journal: iScience

Article Title: Cardiomyocyte-derived HSPB1 regulates TGF-β1 maturation and inhibits endothelial-to-mesenchymal transition in myocardial fibrosis

doi: 10.1016/j.isci.2026.115028

Figure Lengend Snippet: Proposed model of HSPB1-mediated redox regulation of TGF-β1 maturation during post-MI fibrosis. During myocardial fibrosis following myocardial infarction, the expression of HSPB1 is markedly upregulated in the peri-infarct region. Upon activation, HSPB1 exposes its reactive cysteine residue (Cys137), which may interact with critical cysteine sites within pre-pro-TGF-β1, thereby influencing its redox-dependent folding and disulfide bond formation. This interaction potentially interferes with the maturation and secretion of active TGF-β1 into the extracellular space. Reduced secretion of mature TGF-β1 limits Smad2/3 phosphorylation and endothelial-to-mesenchymal transition, ultimately alleviating myocardial fibrosis. The red dashed box highlights the hypothesized redox regulatory interaction between HSPB1 and pre-pro-TGF-β1, which requires further biochemical validation.

Article Snippet: Human umbilical vein endothelial cells (HUVECs; primary, pooled donors; ATCC) were purchased through an authorized distributor in China and originally sourced from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Activation Assay, Residue, Phospho-proteomics, Biomarker Discovery

Journal: iScience

Article Title: Cardiomyocyte-derived HSPB1 regulates TGF-β1 maturation and inhibits endothelial-to-mesenchymal transition in myocardial fibrosis

doi: 10.1016/j.isci.2026.115028

Figure Lengend Snippet: Regulatory role of HSPB1 in endothelial cell EndoMT (A) Western blot shows HSPB1 expression in HUVECs following lentiviral-mediated overexpression (LV-HSPB1) or knockdown (LV-HSPB1-RNAi); β-actin served as a loading control. (B) Quantification of HSPB1/β-actin ratio shows significant differences between groups. (C) Representative images of Transwell migration assays evaluating the effect of HSPB1 on TGF-β1–induced endothelial migration (scale bars, 100 μm). (D) Quantification of migrated cells per field. (E) Representative tube formation images showing the effect of HSPB1 modulation on TGF-β1–induced angiogenic activity (scale bars, 200 μm). (F–H) Quantitative analysis of tube formation parameters, including the number of branches (F), loops (G), and total tube length (H), measured using ImageJ software. Data are presented as mean ± SD ( n ≥ 6). Exact p values are indicated in the graphs. Statistical analyses were performed using one-way ANOVA followed by a Bonferroni post hoc test.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) , ATCC , Primary cells, pooled donors.

Techniques: Western Blot, Expressing, Over Expression, Knockdown, Control, Migration, Activity Assay, Software

Journal: iScience

Article Title: Cardiomyocyte-derived HSPB1 regulates TGF-β1 maturation and inhibits endothelial-to-mesenchymal transition in myocardial fibrosis

doi: 10.1016/j.isci.2026.115028

Figure Lengend Snippet: Effects of HSPB1 on signaling pathways and TGF-β secretion in HUVECs under hypoxic conditions (A and B) HUVECs were transfected with adenoviral vectors for HSPB1 overexpression (OE) or knockdown (KD) and cultured for 48 h before RNA extraction. Gene expression analysis was performed using RNA sequencing. Gene set enrichment analysis (GSEA) assessed the regulatory roles of HSPB1 in processes such as heart development, angiogenesis, and cell proliferation (A). Further analysis using Hallmark gene sets explored HSPB1 signaling pathway activation (B). (C–G) Following transfection, HUVECs were cultured for 24 h and subjected to hypoxic conditions (3% O 2 ) for 48 h. Western blot analysis of the indicated proteins was performed. (D) pSmad2/3/Smad2/3 ratio, (E) quantification of CD31 protein expression, (F) quantification of E-cadherin expression, (G) quantification of α-SMA expression, and (H) quantification of N-cadherin expression were measured relative to β-actin. (I) TGF-β levels were measured by ELISA in cell supernatants. Data are presented as mean ± SD ( n ≥ 6). Exact p values are indicated in the graphs. Statistical analyses were performed using one-way ANOVA followed by a Bonferroni post hoc test.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) , ATCC , Primary cells, pooled donors.

Techniques: Protein-Protein interactions, Transfection, Over Expression, Knockdown, Cell Culture, RNA Extraction, Gene Expression, RNA Sequencing, Activation Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLOS One

Article Title: Glucocorticoids suppress early lung inflammation and impair control of SARS-CoV-2 in non-human primates

doi: 10.1371/journal.pone.0342849

Figure Lengend Snippet: A) Experimental design and sample collection. Created in part with BioRender under CC BY license, Nelson, C., (2025). B) Quantification of total lung lesions detected with 18 FDG-PET/CT imaging at the peak of the response, per animal and per group with mean. Animal #ID represented by color and shape in legend. C) Example 18 FDG-PET/CT images from control animal #DGT7 and GC treated animal #DHXD. D) Quantification of the lung lesion volume (dot size) and the metabolic intensity (normalized FDG uptake = SUV of lesion/SUV muscle). Significance calculated with 2way Anova multiple comparison test at the indicated timepoints between GC treatment and control. E) Quantification of subgenomic RNA of the SARS-CoV-2 N1 protein in copies per mL of BAL fluid. Individual animals and the mean of each group with standard error mean represented. Significance calculated with 2way Anova. Fold change between GC treatment and control indicated. Limit of detection (LOD) is 2,000 copies/mL fluid. F) Subgenomic N1 in copies per gram of tissue in the pulmonary lymph nodes, PET+ involved lung, and PET- uninvolved lung. Significance calculated with 2way Anova multiple comparison test. LOD is 1,000 copies per gram of tissue. p > 0.05 not shown, *p < 0.05, ***p < 0.001.

Article Snippet: Peptide pool consisted of Peptivator SARS-CoV-2 Prot_S Complete (Miltenyi Cat#130-127-953), Peptivator SARS-CoV-2 Prot_S B.1.617.2 Mutation (Miltenyi Cat#130-128-763), and Peptivator SARS-CoV-2 Prot_N (Miltenyi Cat# 130-126-699).

Techniques: Positron Emission Tomography-Computed Tomography, Imaging, Control, Comparison

Journal: PLOS One

Article Title: Glucocorticoids suppress early lung inflammation and impair control of SARS-CoV-2 in non-human primates

doi: 10.1371/journal.pone.0342849

Figure Lengend Snippet: A) Representative flow cytometry of SARS-CoV-2 specific CD4 and CD8 T cell responses (IFNγ + /TNF+) in blood at day 7 post-infection after ex vivo peptide stimulation with Spike and Nucleocapsid peptide pools, gated on live/CD45 + /CD3 + /CD95 + . B) Quantification of the frequency of SARS-CoV-2 specific CD4 and CD8 T cell responses in the blood overtime after subtracting background staining in unstimulated samples. Significance calculated with 2-way Anova and multiple comparison test. C) Representative flow cytometry of SARS-CoV-2 specific CD4 and CD8 T cell responses (IFNγ + /TNF+) in BAL at day 13 post-infection after ex vivo peptide stimulation with Spike and Nucleocapsid peptide pools, gated on live/CD45 + /CD3 + /CD95 + . D) Quantification of the frequency of SARS-CoV-2 specific CD4 and CD8 T cell responses in the BAL overtime after subtracting background staining in unstimulated samples. Significance calculated with 2-way Anova and multiple comparison test. E) Geometric mean fluorescence intensity (GMFI) of IFNγ production by IFNγ + SARS-CoV-2 specific CD4 and CD8 T cells. F) GMFI of TNF production by TNF+ SARS-CoV-2 specific CD4 and CD8 T cells. G) Frequency of IL-2+ of SARS-CoV-2 specific (IFNγ + /TNF+) CD4 and CD8 T cells in the BAL at day 13 post-infection. Significance calculated with unpaired t-test. H) Frequency of Granzyme B+ of SARS-CoV-2 specific (IFNγ + /TNF+) CD4 and CD8 T cells in the BAL at day 13 post-infection. I) Frequency of IL-17A+ of SARS-CoV-2 specific (IFNγ + /TNF+) CD4 and CD8 T cells in the BAL at day 13 post-infection. p > 0.05 ns or not shown, *p < 0.05, ***p < 0.001. ****p < 0.000.

Article Snippet: Peptide pool consisted of Peptivator SARS-CoV-2 Prot_S Complete (Miltenyi Cat#130-127-953), Peptivator SARS-CoV-2 Prot_S B.1.617.2 Mutation (Miltenyi Cat#130-128-763), and Peptivator SARS-CoV-2 Prot_N (Miltenyi Cat# 130-126-699).

Techniques: Flow Cytometry, Infection, Ex Vivo, Staining, Comparison, Fluorescence

Journal: PLOS One

Article Title: Glucocorticoids suppress early lung inflammation and impair control of SARS-CoV-2 in non-human primates

doi: 10.1371/journal.pone.0342849

Figure Lengend Snippet: A) Quantification of total B cells (CD3 - /CD20 + ) by flow cytometry in the blood over time. Mean of each group with standard error mean (SEM) represented. Significance calculated with 2way Anova. B) Representative flow cytometry plots and gating strategy for identifying total B cells (CD3 - /CD20 + ) from the BAL at day 4 after infection. C) Quantification of total B cells by flow cytometry in the BAL over time. Mean, SEM, and 2way Anova. D) Quantification of total B cells by scRNAseq (cluster 9) in the BAL over time. Mean, SEM, and 2way Anova. E) Differentially expressed genes in B cells (cluster 9) at day 4 post-infection between GC treatment vs. control. Red is upregulated with GC treatment, log 2 FC > 1 and adjusted p-value <0.05. Blue is downregulated with GC treatment, log 2 FC < −1 and adjusted p-value <0.05. Grey with large dot is adjusted p-value <0.05 but absolute |log 2 FC| < 1. Grey with small dot is adjusted p-value >0.05. F) Representative flow cytometry plots and gating strategy for identifying naïve B cells (CD3 - /CD20 + /IgD + ) and activated B cell (CD3 - /CD20 + /IgD - /CD95 + ) from the BAL at day 4 after infection. G) Quantification of naïve B cells by flow cytometry in the BAL over time. Mean, SEM, and 2way Anova. H) Quantification of activated B cells by flow cytometry in the BAL over time. Mean, SEM, and 2way Anova. I) Quantification of total B cells, J) naïve B cells, K) and activated B cells in the spleen, bone marrow, axillary lymph nodes, and pulmonary lymph nodes at necropsy day 13 post-infection. Significance calculated with 2way Anova. L) Representative flow cytometry plots of Spike-specific B cells (CD3 - /CD20 + /CD95 + /Spike-tetramer + ) isolated from the axillary or pulmonary lymph nodes at necropsy, day 13 post-infection. M) Quantification of Spike-specific B cells (CD3 - /CD20 + /CD95 + /Spike (B.1.617.2) tetramer + ) as a percentage of non-naïve (IgD - ) B cells in the spleen, bone marrow, axillary lymph nodes, and pulmonary lymph nodes at necropsy day 13 post-infection. N) Quantification of the frequency of IgG + expression by Spike-specific (tetramer + ) B cells in the pulmonary lymph node. Significance calculated with 2way Anova. p > 0.05 ns or not shown, *p < 0.05, ***p < 0.001. ****p < 0.0001.

Article Snippet: Peptide pool consisted of Peptivator SARS-CoV-2 Prot_S Complete (Miltenyi Cat#130-127-953), Peptivator SARS-CoV-2 Prot_S B.1.617.2 Mutation (Miltenyi Cat#130-128-763), and Peptivator SARS-CoV-2 Prot_N (Miltenyi Cat# 130-126-699).

Techniques: Flow Cytometry, Infection, Control, Isolation, Expressing

Journal: Frontiers in Immunology

Article Title: Carbohydrate fatty acid monosulphate ester adjuvant enhances the immunogenicity of influenza antigens via TLR4/2-dependent mechanisms

doi: 10.3389/fimmu.2026.1787181

Figure Lengend Snippet: CMS adjuvant enhances antigen-specific T cell responses to H1N1 HA peptide pools in human PBMCs. (A) Scheme of the AIM assay setup. PBMCs from healthy donors were treated with H1N1 HA peptide pools alone or in combination with the adjuvant for 6 days and restimulated with peptide pools for 16 hrs in the presence of anti-CD40 and anti-CD28, followed by surface staining and flow cytometry analysis of AIMs. (B) Representative dot plots showing frequencies of CD154 + CD137 + CD4 + and CD137 + CD8 + T cells for the indicated treatments. (C) Summarized frequencies of HA-specific CD154 + CD137 + CD4 + and CD137 + CD8 + T cells from independent donors (n=9). Statistics are **P < 0.01 ; ns, not significant as determined by Mann-Whitney’s U test (C, D) . AIM, activation-induced marker; SD, standard deviation.

Article Snippet: The cells were treated with H1N1 HA peptide pools (Miltenyi Biotec, 130-099-803) at a concentration of 0.5 μg/ml alone or combined with CMS (125 μg/ml) for 6 days.

Techniques: Adjuvant, Staining, Flow Cytometry, Activation Assay, Marker, Standard Deviation