primary antibodies  (Bioss)

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: Bulk RNA-seq identification of early regulatory genes and signals in bone defects. ( A ) Experimental workflow for bulk RNA-seq in early stages of bone defects in rats. ( B ) Construction of interaction networks and molecular dynamics simulations for key regulatory proteins in early-stage femoral and alveolar bone defects. ( C – E ) Immunohistochemical detection and quantitative analysis of Heat Shock Protein Family A (Hsp70) Member 5 (HSPA5) and interleukin-6 (IL-6) expression levels in early healing tissues of the femoral and alveolar regions. Scale bars: 2 mm, 100 μm, and 50 μm. ( F , G ) Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis results for differentially expressed genes (DEGs) in early healing stages of femoral and alveolar bone defects. Statistical significance was determined using a t -test.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: RNA Sequencing, Immunohistochemical staining, Expressing

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: Protein identification for key proteins and signals in early bone defects. ( A ) Workflow for tissue protein identification at early stages of bone defects in rats. ( B , C ) KEGG and GO enrichment analysis of differential proteins in early healing tissues of femoral and alveolar bone defects. ( D , E ) Interaction network analysis of HSPA5 and IL-6 within the enrichment signals of femoral and alveolar bones. ( F ) Protein expression levels of HSPA5 and IL-6 in early healing tissues of femoral and alveolar defects. ( G ) Quantitative analysis of HSPA5 and IL-6 protein expression. Statistical analysis employed a t -test.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Expressing

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: IL-6-mediated regulation of HSPA5 in early bone defect repair tissues. ( A ) Extraction of femoral and alveolar bone cell clusters from whole-cell atlases. ( B ) Pseudo-time series analysis of cellular and gene expression in repair tissues. ( C ) The rat model for femoral and alveolar bone defect treatment using HA15 and LMT28. ( D , E ) Immunohistochemical analysis of HSPA5 and IL-6 expression in femoral defects after HA15 treatment. Scale bars: 200 μm and 50 μm. ( F , G ) Immunohistochemical analysis in alveolar defects after LMT28 treatment. Scale bars: 200 μm and 50 μm. HA15 inhibits HSPA5; LMT28 inhibits IL-6. Statistical significance was assessed with a t -test.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Extraction, Gene Expression, Immunohistochemical staining, Expressing

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: M2 macrophages release IL-6 in alveolar bone defect repair. ( A ) Extraction of Il-6 -expressing cell populations. ( B ) Identification and classification of subpopulations within Il-6 -expressing cells. ( C ) Identification of the Il-6 + M2 cell subpopulation. ( D ) Temporal association between Il-6 + M2 cell subpopulation and samples. ( E – G ) Multiplex immunofluorescence for HSPA5 (GRP78/BiP), M2 macrophages (ARG1), and IL-6, combined with statistical analysis of fluorescence intensity and colocalization analysis. Scale bars: 1 mm, 100 μm and 50 μm. ( H ) KEGG and GO enrichment analysis of DEGs in the Il-6 + M2 cell subpopulation.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Extraction, Expressing, Multiplex Assay, Immunofluorescence, Fluorescence

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: IL-6 modulates HSPA5 to mitigate endoplasmic reticulum stress (ERS)-related apoptosis in early bone defects. ( A , B ) Expression and quantitative analysis of ERS-related proteins following HA15 and LMT28 treatments. Scale bars: 200 μm and 50 μm. ( C , D ) Quantitative analysis of apoptosis-related proteins CHOP and caspase-12 after treatment. Scale bars: 200 μm and 50 μm. ( E ) Expression levels of ERS and apoptosis-related proteins were analyzed by Western blot, and statistical analysis was conducted on the results. ( F , G ) Quantitative terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining and statistical analysis of apoptosis in femoral and alveolar bone defects. Scale bars: 100 μm. ( H ) Ca 2+ histopathological staining. Scale bars: 100 μm and 40 μm. ( I ) Detection of reactive oxygen species in repair tissue following HA15 and LMT28 treatment. Scale bars: 200 μm and 50 μm. Statistical significance was determined using one-way ANOVA.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Expressing, Western Blot, TUNEL Assay, Staining

Journal: Veterinary Research

Article Title: Outer membrane vesicles secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing endoplasmic reticulum stress-mediated autophagy flux blockade

doi: 10.1186/s13567-025-01679-6

Figure Lengend Snippet: APEC OMVs induce ERS in HD11 cells to evade immune clearance by macrophages . Transmission electron microscopy shows ER network expansion and swelling in HD11 cells treated with WT, WTΔ ypjA , or OMVs. The data represent one of three independent experiments. Scale bar, 2 μm. B OMVs colocalized with ER. DiO-OMVs were incubated with HD11 cells at 37 °C for 6 h, the ER was labeled with ER-Tracker Red, and laser confocal microscopy imaging was performed. The data represent one of three independent experiments. Scale bar, 7.5 μm. Colocalization scatter plot was generated using ImageJ software and GraphPad Prism 8. C HD11 cells were treated with 50–200 µg/mL OMVs for 6 h, and the relative mRNA level of GRP78/BiP was determined by qPCR. ( n = 3) D HD11 cells were treated with OMVs (100 µg/mL) for 0–9 h, and the relative mRNA level of GRP78/BiP was determined by qPCR. ( n = 3) E , F HD11 cells were treated with OMVs (100 µg/mL) for 0–9 h, and western blot was used to detect the expression of GRP78/BiP protein in cell lysates and gray value analysis was performed using ImageJ software ( F ). ( n = 3) G Cell viability of HD11 cells treated with OMVs (200 µg/mL) for 12 h was detected by CCK-8 in the absence or presence of 4-PBA (2 mM). ( n = 3) H , I Intracellular survival of HD11 cells infected with WT at MOI = 100 in the absence or presence of 4-PBA (2 mM). Square culture plates showed 1/ 100 of the bacterial load. ( n = 3). n represents three biological replicates; data points indicate independent culture systems. Bar charts display mean ± standard error of the mean (SEM). Data were analyzed using Student’s t -test and two-way ANOVA with Sidak correction. (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Rabbit anti-GRP78/BiP polyclonal antibody , Abmart, Shanghai, China , 1:5000.

Techniques: Transmission Assay, Electron Microscopy, Incubation, Labeling, Confocal Microscopy, Imaging, Generated, Software, Western Blot, Expressing, CCK-8 Assay, Infection

Journal: Veterinary Research

Article Title: Outer membrane vesicles secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing endoplasmic reticulum stress-mediated autophagy flux blockade

doi: 10.1186/s13567-025-01679-6

Figure Lengend Snippet: APEC OMVs induce ERS to promote APEC systemic infection . Flowchart of the infection experiment using chicks treated with 4-PBA. B On the sixth day after WT infection of chicks (1 × 10 CFU/chick), the number of bacteria (CFU) was counted in the trachea, lungs, liver, and spleen tissues of the chicks in the absence or presence of 4-PBA (50 mg/kg). Trachea: 95% CI [0.26–0.38]; lungs: 95% CI [0.65–0.78]; liver: 95% CI [0.004–0.13]; spleen: 95% CI [0.08–0.21]. C On the sixth day after WT infection of chicks (1 × 10 9 CFU/chick), HE staining was performed on tracheal, lung, liver, and spleen tissues of the chicks in the absence or presence of 4-PBA (50 mg/kg). Trachea: degeneration and hyperplasia of epithelial mucosal cells (black arrow), diffuse neutrophilic infiltration (red arrow), mucus and inflammatory exudate in the mucosa of the tracheal wall (blue arrow); lung: markedly widened pulmonary septa (black arrow), inflammatory cell infiltration in the pulmonary interstitium (red arrow); liver: hepatocytes show lytic degeneration (black arrow) with inflammatory cell infiltration (red arrow); spleen: marked hyperplasia of lymphoid follicles (black arrow), blurred demarcation between white and red pulp (red arrow). Scale bar, 50 µm or 200 µm. D – F GRP78/BiP immunofluorescence staining was performed on lung and spleen tissues from chicks on day 6 post-infection ( D ) and analyzed using ImageJ software ( E , F ). Scale bar, 50 µm. Data points represent independent cultures; bar charts show mean ± standard error of the mean (SEM). Two-way ANOVA with Sidak correction was performed (* p < 0.05, ** p < 0.01, *** p < 0.001). Effect sizes with 95% confidence intervals are reported in panel ( B ).

Article Snippet: Rabbit anti-GRP78/BiP polyclonal antibody , Abmart, Shanghai, China , 1:5000.

Techniques: Infection, Bacteria, Staining, Immunofluorescence, Software

Journal: Veterinary Research

Article Title: Outer membrane vesicles secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing endoplasmic reticulum stress-mediated autophagy flux blockade

doi: 10.1186/s13567-025-01679-6

Figure Lengend Snippet: OMVs secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing ERS-mediated autophagy flux blockade . APEC-secreted OMVs, upon uptake by HD11 cells, induce ROS accumulation and Ca 2+ release, triggering ERS and activating UPR pathways, including the PERK, IRE1, and ATF6 signaling branches, leading to the expression of stress-related factors such as GRP78/BiP and CHOP. The sustained activation of ERS inhibits autophagosome degradation and disrupts the acidic environment of lysosomes, thereby preventing autophagosomes from fusing with lysosomes and impairing the phagocytic clearance capacity of macrophages. Collectively, these abnormal conditions facilitate APEC survival within HD11 cells and enable immune evasion, ultimately promoting bacterial dissemination and systemic infection in the host.

Article Snippet: Rabbit anti-GRP78/BiP polyclonal antibody , Abmart, Shanghai, China , 1:5000.

Techniques: Infection, Expressing, Activation Assay