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: Discover Oncology

Article Title: The influence of ferroptosis-associated gene HSPA5 on the prognosis and immune evasion of lung adenocarcinoma

doi: 10.1007/s12672-025-03629-2

Figure Lengend Snippet: DEGs of ferroptosis in lung adenocarcinoma. A Volcano map revealed DEGs of LUAD in GSE27262 database. B Heat map revealed DEGs of LUAD in GSE27262 database. C Volcano map revealed DEGs of LUAD in TCGA database. D Heat map revealed DEGs of LUAD in TCGA database. E Venn diagram of GSE27262 database and TCGA database. F Expression of ACSL4, DPP4, HSPA5 and HSPB1 in the GSE27262 database. G Expression of ACSL4, DPP4, HSPA5 and HSPB1 in the TCGA database

Article Snippet: The GAPDH antibody purchased from Cell Signaling Technology (Danvers, MA, USA) was utilized at a dilution ratio of 1:1000 (product code: 5174 S), with the exposure time of 5 s. Additionally, the Anti-HSPA5 antibody was acquired from Proteintech (Rosemont, PA, USA), and utilized at a dilution ratio of 1:1000 (product code: 66574-1-Ig), with the exposure time of 10 s.

Techniques: Expressing

Journal: Discover Oncology

Article Title: The influence of ferroptosis-associated gene HSPA5 on the prognosis and immune evasion of lung adenocarcinoma

doi: 10.1007/s12672-025-03629-2

Figure Lengend Snippet: Prognostic analysis of ferroptosis genes in lung adenocarcinoma. A Upset plot revealed Prognostic of LUAD in TCGA, GSE31210 , and GSE26939 databases. B Survival probability of HSPA5, ACSL4, DPP4, HSPB1 in TCGA, GSE31210 and GSE26939 databases

Article Snippet: The GAPDH antibody purchased from Cell Signaling Technology (Danvers, MA, USA) was utilized at a dilution ratio of 1:1000 (product code: 5174 S), with the exposure time of 5 s. Additionally, the Anti-HSPA5 antibody was acquired from Proteintech (Rosemont, PA, USA), and utilized at a dilution ratio of 1:1000 (product code: 66574-1-Ig), with the exposure time of 10 s.

Techniques:

Journal: Discover Oncology

Article Title: The influence of ferroptosis-associated gene HSPA5 on the prognosis and immune evasion of lung adenocarcinoma

doi: 10.1007/s12672-025-03629-2

Figure Lengend Snippet: Prediction of ferroptosis gene regulation. A Screening of candidate genes. B Association of HSPA5 with transcription factors. (X-axes: the expression of transcription factors; Y-axes: the expression of HSPA5). C CeRNA network of ferroptosis genes

Article Snippet: The GAPDH antibody purchased from Cell Signaling Technology (Danvers, MA, USA) was utilized at a dilution ratio of 1:1000 (product code: 5174 S), with the exposure time of 5 s. Additionally, the Anti-HSPA5 antibody was acquired from Proteintech (Rosemont, PA, USA), and utilized at a dilution ratio of 1:1000 (product code: 66574-1-Ig), with the exposure time of 10 s.

Techniques: Expressing

Journal: Discover Oncology

Article Title: The influence of ferroptosis-associated gene HSPA5 on the prognosis and immune evasion of lung adenocarcinoma

doi: 10.1007/s12672-025-03629-2

Figure Lengend Snippet: Ferroptosis gene DNA methylation regulation and comprehensive regulatory network. A RNA methylation regulation of Ferroptosis gene. B Regulatory sites of DNA methylation and RNA methylation for ALOX15, FANCD2, HSPA5, RPL8, SLC1A5 and SLC7A11. C Integrated regulatory network of ALOX15, FANCD2, HSPA5, RPL8, SLC1A5 and SLC7A11

Article Snippet: The GAPDH antibody purchased from Cell Signaling Technology (Danvers, MA, USA) was utilized at a dilution ratio of 1:1000 (product code: 5174 S), with the exposure time of 5 s. Additionally, the Anti-HSPA5 antibody was acquired from Proteintech (Rosemont, PA, USA), and utilized at a dilution ratio of 1:1000 (product code: 66574-1-Ig), with the exposure time of 10 s.

Techniques: DNA Methylation Assay, Methylation

Journal: Discover Oncology

Article Title: The influence of ferroptosis-associated gene HSPA5 on the prognosis and immune evasion of lung adenocarcinoma

doi: 10.1007/s12672-025-03629-2

Figure Lengend Snippet: HSPA5 regulates LUAD cell proliferation, migration and cell apoptosis in vitro. A HSPA5 expression levels were analyzed after transfection of siRNA compared with the control groups in A549 and PC-9 cells by Western blotting. B MTT assays were used to detect the viability of A549 and PC-9 cells after HSPA5 was silenced compared with the control groups. C After knocking down HSPA5, the colony formation abilities of A549 and PC-9 cells were detected. D The proliferation of A549 and PC-9 cells was analyzed by EdU after transfection. E Transwell assays were performed to detect the migration ability of A549 and PC-9 cells after HSPA5 knockdown. F A549 cells was stained and analyzed by flow cytometry. G The detection of ROS level with corresponding treatment in A549 and PC-9 cells. H The detection of iron level with corresponding treatment in A549 and PC-9 cells. * p < 0.05, ** p < 0.01

Article Snippet: The GAPDH antibody purchased from Cell Signaling Technology (Danvers, MA, USA) was utilized at a dilution ratio of 1:1000 (product code: 5174 S), with the exposure time of 5 s. Additionally, the Anti-HSPA5 antibody was acquired from Proteintech (Rosemont, PA, USA), and utilized at a dilution ratio of 1:1000 (product code: 66574-1-Ig), with the exposure time of 10 s.

Techniques: Migration, In Vitro, Expressing, Transfection, Control, Western Blot, Knockdown, Staining, Flow Cytometry