hspa5  (R&D Systems)

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: Cell Death & Disease

Article Title: HSPA5 promotes YAP/TAZ stability independently of the Hippo pathway and induces proneural-to-mesenchymal transition in glioblastoma

doi: 10.1038/s41419-026-08428-3

Figure Lengend Snippet: A Heatmap illustrating the mRNA expression levels of HSP70 family genes across Proneural (PN), Classical (CL), and Mesenchymal (MES) glioblastoma (GBM) subtypes from TCGA datasets. B Heatmap of HSP70 family gene expression in PN, CL, and MES subtypes from the Gulou GBM datasets. C , D The expression levels of HSPA5 ( C ) and HSPA6 ( D ) in normal brain tissues (NBTs) and GBM subtypes from TCGA dataset. E , F The expression levels of HSPA5 ( E ) and HSPA6 ( F ) in PN, CL, and MES GBM subtypes from the Gulou GBM datasets. G Representative immunohistochemical (IHC) staining images of HSPA5 protein in NBTs, PN, CL, and MES GBM samples. Scale bar: 50 μm. H , I Kaplan–Meier survival curves for GBM patients stratified by median HSPA5 ( H ) and HSPA6 ( I ) expression levels in TCGA cohort. P values were calculated using the log-rank test. J GSEA showed a significant positive correlation between the expression of HSPA5 and MES subtypes, and a negative correlation with PN subtypes. ns: P > 0.05, * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: The primary antibodies used for western blot analysis were as follows: HSPA5 (1:1000, Cat No. 66574-1-Ig, Proteintech, China), HSPA5 (1:1000, Cat No. 11587-1-AP, Proteintech, China), YAP (1:1000, Cat No. 13584-1-AP, Proteintech, China), TAZ (1:1000, Cat No. 23306-1-AP, Proteintech, China), p-YAP (S127) (1:1000, Cat No. 4911, Cell Signaling Technology, USA), p-TAZ (S89) (1:1000, Cat No. 59971, Cell Signaling Technology, USA), β-TrCP (1:1000, Cat No. 28393-1-AP, Proteintech, China), c-MET (1:1000, Cat No. 25869-1-AP, Proteintech, China), CD44 (1:1000, Cat No. 15675-1-AP, Proteintech, China), SOX2 (1:1000, Cat No. 11064-1-AP, Proteintech, China), OLIG2 (1:1000, Cat No. 13999-1-AP, Proteintech, China), Ub (1:2000, Cat No. A19686, Abclonal, China), GAPDH (1:5000, Cat No. 60004-1-Ig, Proteintech, China), GAPDH (1:5000, Cat No. 10494-1-AP, Proteintech, China), H3 (1:2000, Cat No. 17168-1-AP, Proteintech, China), GST (1:1000, Cat No. 10000-0-AP, Proteintech, China), Flag (1:1000, Cat No. 20543-1-AP, Proteintech, China), His (1:1000, Cat No. 66005-1-Ig, Proteintech, China), Myc (1:1000, Cat No. 60003-2-Ig, Proteintech, China).

Techniques: Expressing, Gene Expression, Immunohistochemical staining, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: HSPA5 promotes YAP/TAZ stability independently of the Hippo pathway and induces proneural-to-mesenchymal transition in glioblastoma

doi: 10.1038/s41419-026-08428-3

Figure Lengend Snippet: A Primary GBM cells (PN20, PN24, MES50 and MES52) were isolated from post-surgical GBM tissues. B Western blot (WB) analysis of HSPA5, OLIG2, SOX2, c-MET, and CD44 protein expression in NHA, PN (PN20 and PN24), and MES (MES50 and MES52) cells. C Representative IHC staining of OLIG2, SOX2, CD44, and c-MET in clinical PN and MES GBM specimens. Scale bar: 20 μm. D , E CCK-8 assay was used to analyze the cell viability of MES cells transduced with shNC and shHSPA5. F , G CCK-8 assay was used to analyze the cell viability of PN cells transduced with Vector and HSPA5. H , I Colony formation assay was used to analyze the cell proliferation ability of MES cells transduced with shNC and shHSPA5. J , K Colony formation assay was used to analyze the cell proliferation ability of PN cells transduced with Vector and HSPA5. L Spearman correlation analysis between HSPA5 expression and PN ( OLIG2 and SOX2 ) or MES ( CD44 and c-MET ) subtype biomarkers in TCGA GBM datasets. M WB analysis of HSPA5, OLIG2, SOX2, c-MET, CD44, C/EBPβ, N-cadherin, STAT3, and p-STAT3 protein levels in MES cells following HSPA5 knockdown. N WB analysis of HSPA5, OLIG2, SOX2, c-MET, CD44, C/EBPβ, N-cadherin, STAT3, and p-STAT3 protein levels in PN cells following HSPA5 overexpression. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 and *** P < 0.001. Two-tailed unpaired t test ( D – K ).

Article Snippet: The primary antibodies used for western blot analysis were as follows: HSPA5 (1:1000, Cat No. 66574-1-Ig, Proteintech, China), HSPA5 (1:1000, Cat No. 11587-1-AP, Proteintech, China), YAP (1:1000, Cat No. 13584-1-AP, Proteintech, China), TAZ (1:1000, Cat No. 23306-1-AP, Proteintech, China), p-YAP (S127) (1:1000, Cat No. 4911, Cell Signaling Technology, USA), p-TAZ (S89) (1:1000, Cat No. 59971, Cell Signaling Technology, USA), β-TrCP (1:1000, Cat No. 28393-1-AP, Proteintech, China), c-MET (1:1000, Cat No. 25869-1-AP, Proteintech, China), CD44 (1:1000, Cat No. 15675-1-AP, Proteintech, China), SOX2 (1:1000, Cat No. 11064-1-AP, Proteintech, China), OLIG2 (1:1000, Cat No. 13999-1-AP, Proteintech, China), Ub (1:2000, Cat No. A19686, Abclonal, China), GAPDH (1:5000, Cat No. 60004-1-Ig, Proteintech, China), GAPDH (1:5000, Cat No. 10494-1-AP, Proteintech, China), H3 (1:2000, Cat No. 17168-1-AP, Proteintech, China), GST (1:1000, Cat No. 10000-0-AP, Proteintech, China), Flag (1:1000, Cat No. 20543-1-AP, Proteintech, China), His (1:1000, Cat No. 66005-1-Ig, Proteintech, China), Myc (1:1000, Cat No. 60003-2-Ig, Proteintech, China).

Techniques: Isolation, Western Blot, Expressing, Immunohistochemistry, CCK-8 Assay, Transduction, Plasmid Preparation, Colony Assay, Knockdown, Over Expression, Two Tailed Test

Journal: Cell Death & Disease

Article Title: HSPA5 promotes YAP/TAZ stability independently of the Hippo pathway and induces proneural-to-mesenchymal transition in glioblastoma

doi: 10.1038/s41419-026-08428-3

Figure Lengend Snippet: A The RNA-seq results were used to identify the enriched pathways after knockdown of HSPA5 expression in MES50 cells. B A schematic illustration of the Hippo pathway in mammals. The activated LATS1/2 phosphorylated and inactivated YAP/TAZ, preventing it from translocating into the nucleus and binding to transcription factors TEAD1–4. C Heatmap showing expression levels of YAP/TAZ downstream target genes in MES50-shNC and MES50-shHSPA5 cells. D Effect of HSPA5 knockdown (shNC, shHSPA5#1, and shHSPA5#2) on 8xGTIIC-Luc activity in HEK293T cells transfected with YAP or TAZ expression vectors. E Effect of HSPA5 overexpression (Vector and HSPA5) on 8xGTIIC-Luc activity in HEK293T cells transfected with YAP or TAZ expression vectors. F Effect of HSPA5 knockdown (shNC, shHSPA5#1, and shHSPA5#2) on CTGF Luc reporter transcriptional activity in HEK293T cells transfected with YAP or TAZ expression vectors. G Effect of HSPA5 overexpression (Vector and HSPA5) on CTGF Luc reporter transcriptional activity in HEK293T cells transfected with YAP or TAZ expression vectors. H , I qRT-PCR analysis of the expression of YAP/TAZ downstream target genes ( CTGF and CYR61 ) in MES cells transduced with shNC and shHSPA5. J , K qRT-PCR analysis of the expression of YAP/TAZ downstream target genes ( CTGF and CYR61 ) in PN cells transduced with Vector and HSPA5. L WB analysis of protein expression of HSPA5, OLIG2, SOX2, c-MET, CD44, C/EBPβ, N-cadherin, STAT3, and p-STAT3 in shNC/shHSPA5 MES cells treated with indicated interventions (transfection of unphosphorylatable YAP-5SA/TAZ-4SA). M WB analysis of protein expression of HSPA5, OLIG2, SOX2, c-MET, CD44, C/EBPβ, N-cadherin, STAT3, and p-STAT3 in Vector/HSPA5 PN cells treated with indicated interventions (knockdown of YAP/TAZ). Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 and *** P < 0.001. Two-tailed unpaired t test ( D – K ).

Article Snippet: The primary antibodies used for western blot analysis were as follows: HSPA5 (1:1000, Cat No. 66574-1-Ig, Proteintech, China), HSPA5 (1:1000, Cat No. 11587-1-AP, Proteintech, China), YAP (1:1000, Cat No. 13584-1-AP, Proteintech, China), TAZ (1:1000, Cat No. 23306-1-AP, Proteintech, China), p-YAP (S127) (1:1000, Cat No. 4911, Cell Signaling Technology, USA), p-TAZ (S89) (1:1000, Cat No. 59971, Cell Signaling Technology, USA), β-TrCP (1:1000, Cat No. 28393-1-AP, Proteintech, China), c-MET (1:1000, Cat No. 25869-1-AP, Proteintech, China), CD44 (1:1000, Cat No. 15675-1-AP, Proteintech, China), SOX2 (1:1000, Cat No. 11064-1-AP, Proteintech, China), OLIG2 (1:1000, Cat No. 13999-1-AP, Proteintech, China), Ub (1:2000, Cat No. A19686, Abclonal, China), GAPDH (1:5000, Cat No. 60004-1-Ig, Proteintech, China), GAPDH (1:5000, Cat No. 10494-1-AP, Proteintech, China), H3 (1:2000, Cat No. 17168-1-AP, Proteintech, China), GST (1:1000, Cat No. 10000-0-AP, Proteintech, China), Flag (1:1000, Cat No. 20543-1-AP, Proteintech, China), His (1:1000, Cat No. 66005-1-Ig, Proteintech, China), Myc (1:1000, Cat No. 60003-2-Ig, Proteintech, China).

Techniques: RNA Sequencing, Knockdown, Expressing, Binding Assay, Activity Assay, Transfection, Over Expression, Plasmid Preparation, Quantitative RT-PCR, Transduction, Two Tailed Test

Journal: Cell Death & Disease

Article Title: HSPA5 promotes YAP/TAZ stability independently of the Hippo pathway and induces proneural-to-mesenchymal transition in glioblastoma

doi: 10.1038/s41419-026-08428-3

Figure Lengend Snippet: A , B WB analysis of total YAP, TAZ, and their phosphorylated forms (p-YAP S127, and p-TAZ S89) protein levels in MES (A) and PN (B) cells following HSPA5 knockdown or overexpression. C WB analysis of YAP and TAZ protein levels in cytoplasmic and nuclear fractions from indicated cells. GAPDH and Histone H3 serve as fractionation controls. D , E WB analysis of the Hippo pathway components in serum-starved MES ( D ) and PN ( E ) cells following HSPA5 knockdown or overexpression. F WB analysis of YAP/TAZ in control (shNC) and LATS1/2 double-knockdown (shLATS1/2) MES50 cells with or without HSPA5 knockdown. G qRT-PCR analysis of YAP1 and WWTR1 mRNA levels in MES cells transduced with sh NC and sh HSPA5 knockdown. H , I WB analysis of YAP/TAZ protein levels in HSPA5-knockdown MES cells treated with proteasome inhibitor MG132 (20 μM) or DMSO vehicle control for 8 h. J , K WB analysis of total lysates derived from PN20 and MES50 cells stably expressing empty vector, HSPA5, shNC or shHSPA5 and treated with cycloheximide (CHX, 20 μg/mL) for the indicated time points. GAPDH was analyzed as an internal loading control. Quantification of the band intensities normalized to the t = 0 controls is shown in the right panel. L , M WB analysis of total lysates and immunoprecipitates from HSPA5 knockdown MES50 cells. Cells were treated with MG132 (20 µM for 8 h), and the endogenous poly-ubiquitinated YAP ( L ) and TAZ ( M ) were immunoprecipitated with anti-YAP or anti-TAZ antibodies, respectively, and immunoblotted with anti-Ub antibodies. N , O WB analysis of total lysates and immunoprecipitates from HSPA5 overexpression PN20 cells. Cells were treated with MG132 (20 µM for 8 h), and the endogenous poly-ubiquitinated YAP ( N ) and TAZ ( O ) were immunoprecipitated with anti-YAP or anti-TAZ antibodies, respectively, and immunoblotted with anti-Ub antibodies. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 and *** P < 0.001. Two-tailed unpaired t test ( G , J , and K ).

Article Snippet: The primary antibodies used for western blot analysis were as follows: HSPA5 (1:1000, Cat No. 66574-1-Ig, Proteintech, China), HSPA5 (1:1000, Cat No. 11587-1-AP, Proteintech, China), YAP (1:1000, Cat No. 13584-1-AP, Proteintech, China), TAZ (1:1000, Cat No. 23306-1-AP, Proteintech, China), p-YAP (S127) (1:1000, Cat No. 4911, Cell Signaling Technology, USA), p-TAZ (S89) (1:1000, Cat No. 59971, Cell Signaling Technology, USA), β-TrCP (1:1000, Cat No. 28393-1-AP, Proteintech, China), c-MET (1:1000, Cat No. 25869-1-AP, Proteintech, China), CD44 (1:1000, Cat No. 15675-1-AP, Proteintech, China), SOX2 (1:1000, Cat No. 11064-1-AP, Proteintech, China), OLIG2 (1:1000, Cat No. 13999-1-AP, Proteintech, China), Ub (1:2000, Cat No. A19686, Abclonal, China), GAPDH (1:5000, Cat No. 60004-1-Ig, Proteintech, China), GAPDH (1:5000, Cat No. 10494-1-AP, Proteintech, China), H3 (1:2000, Cat No. 17168-1-AP, Proteintech, China), GST (1:1000, Cat No. 10000-0-AP, Proteintech, China), Flag (1:1000, Cat No. 20543-1-AP, Proteintech, China), His (1:1000, Cat No. 66005-1-Ig, Proteintech, China), Myc (1:1000, Cat No. 60003-2-Ig, Proteintech, China).

Techniques: Knockdown, Over Expression, Fractionation, Control, Quantitative RT-PCR, Transduction, Derivative Assay, Stable Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Two Tailed Test

Journal: Cell Death & Disease

Article Title: HSPA5 promotes YAP/TAZ stability independently of the Hippo pathway and induces proneural-to-mesenchymal transition in glioblastoma

doi: 10.1038/s41419-026-08428-3

Figure Lengend Snippet: A WB analysis of total lysates from HEK293T cells co-transfected with FLAG-YAP or FLAG-TAZ, HA-β-TrCP, and HSPA5 expression vectors as indicated. Cells were treated with MG132 (20 µM) for 8 h prior to lysis. B , C WB analysis of total lysates and immunoprecipitates from HEK293T cells stably expressing FLAG-YAP ( B ) or FLAG-TAZ (C), HA-Ub, HA-β-TrCP, and HSPA5. Cells were treated with MG132 (20 µM for 8 h), Flag-tagged proteins were immunoprecipitated, and poly-ubiquitination was detected using anti-HA-Ub antibodies. D WB analysis of protein expression of HSPA5, β-TrCP, YAP and TAZ in HSPA5-depletion MES50 cells and transfected with siNC and siβ-TrCP siRNA. E Schematic workflow for the identification of HSPA5-interacting proteins. Endogenous HSPA5 was immunoprecipitated from MES50 cell lysates, and bound proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). F The Venn diagram showing the overlap (n = 59) between HSPA5-interacting proteins identified by mass spectrometry and predicted interactors from the HitPredict database. G The table presents the rankings and predicted scores of the top 10 peptides, including peptides derived from YAP and TAZ proteins. H , I Endogenous co-immunoprecipitation (Co-IP) in MES50 cells. Cell lysates were immunoprecipitated with anti-YAP ( H ), anti-TAZ ( I ), or anti-HSPA5 ( I ) antibodies, and corresponding IgG controls. Immunoprecipitates and inputs were analyzed by WB. J Schematic representation of the domain structures of full-length and deletion mutants of HSPA5, YAP, and TAZ used in this study. K , L Plasmids containing full-length and truncated HSPA5 constructs were generated, and HEK293T cells were transfected with the indicated plasmids. The Co-IP assay was performed to explore the binding regions between HSPA5 and YAP ( K ) or TAZ ( L ). M , N Plasmids containing full-length and truncated YAP and TAZ constructs were generated, and HEK293T cells were transfected with the indicated plasmids. The Co-IP assay was performed to explore the binding regions between HSPA5 and YAP ( M ) or TAZ ( N ). O , P The ubiquitination assay was conducted using exogenously expressed HA-Ub, His-β-TrCP, FLAG-YAP or FLAG-TAZ, and the indicated HSPA5 truncation mutants. Cells were treated with MG132 (20 µM for 8 h), and immunoprecipitation was performed to assess the ubiquitination of TAZ ( O ) and TAZ ( P ). Q , R Endogenous Co-IP assays were performed in MES50-shNC and MES50-shHSPA5 cells following MG132 (20 µM for 8 h) treatment. The interaction between β-TrCP and YAP ( Q ) or TAZ ( R ) were subsequently examined.

Article Snippet: The primary antibodies used for western blot analysis were as follows: HSPA5 (1:1000, Cat No. 66574-1-Ig, Proteintech, China), HSPA5 (1:1000, Cat No. 11587-1-AP, Proteintech, China), YAP (1:1000, Cat No. 13584-1-AP, Proteintech, China), TAZ (1:1000, Cat No. 23306-1-AP, Proteintech, China), p-YAP (S127) (1:1000, Cat No. 4911, Cell Signaling Technology, USA), p-TAZ (S89) (1:1000, Cat No. 59971, Cell Signaling Technology, USA), β-TrCP (1:1000, Cat No. 28393-1-AP, Proteintech, China), c-MET (1:1000, Cat No. 25869-1-AP, Proteintech, China), CD44 (1:1000, Cat No. 15675-1-AP, Proteintech, China), SOX2 (1:1000, Cat No. 11064-1-AP, Proteintech, China), OLIG2 (1:1000, Cat No. 13999-1-AP, Proteintech, China), Ub (1:2000, Cat No. A19686, Abclonal, China), GAPDH (1:5000, Cat No. 60004-1-Ig, Proteintech, China), GAPDH (1:5000, Cat No. 10494-1-AP, Proteintech, China), H3 (1:2000, Cat No. 17168-1-AP, Proteintech, China), GST (1:1000, Cat No. 10000-0-AP, Proteintech, China), Flag (1:1000, Cat No. 20543-1-AP, Proteintech, China), His (1:1000, Cat No. 66005-1-Ig, Proteintech, China), Myc (1:1000, Cat No. 60003-2-Ig, Proteintech, China).

Techniques: Transfection, Expressing, Lysis, Stable Transfection, Immunoprecipitation, Ubiquitin Proteomics, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Derivative Assay, Co-Immunoprecipitation Assay, Construct, Generated, Binding Assay

Journal: Cell Death & Disease

Article Title: HSPA5 promotes YAP/TAZ stability independently of the Hippo pathway and induces proneural-to-mesenchymal transition in glioblastoma

doi: 10.1038/s41419-026-08428-3

Figure Lengend Snippet: A A schematic of the experimental procedure for establishing the GBM intracranial xenograft mouse model. B – D Bioluminescence images ( B ), quantification of bioluminescence intensity at day 28 ( C ), and Kaplan–Meier survival curves ( D ) of mice implanted with luciferase-expressing MES50 cells expressing shNC, shHSPA5, or shHSPA5 + YAP-5SA/TAZ-4SA. n = 8 mice per group. E Representative H&E-staining images of nice intracranial GBM xenograft models derived from luciferase-expressing MES50 cells transfected with indicated vectors. Scale bar: 50 μm. F – H Bioluminescence images ( F ), quantification bioluminescence intensity at day 28 ( G ), and Kaplan–Meier survival curves ( H ) of mice implanted with luciferase-expressing PN20 cells expressing Vector, HSPA5, and HSPA5+shYAP/TAZ. n = 8 mice per group. I Representative H&E-staining images of nice intracranial GBM xenograft models derived from luciferase-expressing PN20 cells transfected with indicated vectors. Scale bar: 50 μm. J Immunohistochemical staining was used to examine the expression levels of HSPA5, YAP/TAZ, CD44, OLIG2, and Ki-67 in xenograft tumor tissues from different experimental groups. Scale bar: 50 μm. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 and *** P < 0.001. Two-tailed unpaired t test ( C , G ). Log-rank test ( D , H ).

Article Snippet: The primary antibodies used for western blot analysis were as follows: HSPA5 (1:1000, Cat No. 66574-1-Ig, Proteintech, China), HSPA5 (1:1000, Cat No. 11587-1-AP, Proteintech, China), YAP (1:1000, Cat No. 13584-1-AP, Proteintech, China), TAZ (1:1000, Cat No. 23306-1-AP, Proteintech, China), p-YAP (S127) (1:1000, Cat No. 4911, Cell Signaling Technology, USA), p-TAZ (S89) (1:1000, Cat No. 59971, Cell Signaling Technology, USA), β-TrCP (1:1000, Cat No. 28393-1-AP, Proteintech, China), c-MET (1:1000, Cat No. 25869-1-AP, Proteintech, China), CD44 (1:1000, Cat No. 15675-1-AP, Proteintech, China), SOX2 (1:1000, Cat No. 11064-1-AP, Proteintech, China), OLIG2 (1:1000, Cat No. 13999-1-AP, Proteintech, China), Ub (1:2000, Cat No. A19686, Abclonal, China), GAPDH (1:5000, Cat No. 60004-1-Ig, Proteintech, China), GAPDH (1:5000, Cat No. 10494-1-AP, Proteintech, China), H3 (1:2000, Cat No. 17168-1-AP, Proteintech, China), GST (1:1000, Cat No. 10000-0-AP, Proteintech, China), Flag (1:1000, Cat No. 20543-1-AP, Proteintech, China), His (1:1000, Cat No. 66005-1-Ig, Proteintech, China), Myc (1:1000, Cat No. 60003-2-Ig, Proteintech, China).

Techniques: Luciferase, Expressing, Staining, Derivative Assay, Transfection, Plasmid Preparation, Immunohistochemical staining, Two Tailed Test

Journal: Cell Death & Disease

Article Title: HSPA5 promotes YAP/TAZ stability independently of the Hippo pathway and induces proneural-to-mesenchymal transition in glioblastoma

doi: 10.1038/s41419-026-08428-3

Figure Lengend Snippet: A Representative image showing HSPA5, YAP, and TAZ protein expression in two matched pairs of primary (PN subtype) and their corresponding recurrent (MES subtype) GBM specimens. Scale bar: 50 μm. B , C Spearman correlation analysis between HSPA5 and YAP1 or WWTR1 mRNA levels in the Gulou GBM datasets ( n = 50 patient samples). D qRT-PCR analysis of HSPA5 , YAP1 , and WWTR1 mRNA levels in PN and MES GBM specimens from the Gulou cohort. E Kaplan–Meier survival analysis for GBM patients in the Gulou cohort stratified by median HSPA5 , YAP1 , and WWTR1 expression levels in GBM. P values were calculated using the log-rank test. F Schematic illustration of the current study: HSPA5 interacts with and stabilizes YAP/TAZ by disrupting their interaction with β-TrCP, thereby promoting PMT in GBM. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 and *** P < 0.001. Two-tailed unpaired t test ( D – F ).

Article Snippet: The primary antibodies used for western blot analysis were as follows: HSPA5 (1:1000, Cat No. 66574-1-Ig, Proteintech, China), HSPA5 (1:1000, Cat No. 11587-1-AP, Proteintech, China), YAP (1:1000, Cat No. 13584-1-AP, Proteintech, China), TAZ (1:1000, Cat No. 23306-1-AP, Proteintech, China), p-YAP (S127) (1:1000, Cat No. 4911, Cell Signaling Technology, USA), p-TAZ (S89) (1:1000, Cat No. 59971, Cell Signaling Technology, USA), β-TrCP (1:1000, Cat No. 28393-1-AP, Proteintech, China), c-MET (1:1000, Cat No. 25869-1-AP, Proteintech, China), CD44 (1:1000, Cat No. 15675-1-AP, Proteintech, China), SOX2 (1:1000, Cat No. 11064-1-AP, Proteintech, China), OLIG2 (1:1000, Cat No. 13999-1-AP, Proteintech, China), Ub (1:2000, Cat No. A19686, Abclonal, China), GAPDH (1:5000, Cat No. 60004-1-Ig, Proteintech, China), GAPDH (1:5000, Cat No. 10494-1-AP, Proteintech, China), H3 (1:2000, Cat No. 17168-1-AP, Proteintech, China), GST (1:1000, Cat No. 10000-0-AP, Proteintech, China), Flag (1:1000, Cat No. 20543-1-AP, Proteintech, China), His (1:1000, Cat No. 66005-1-Ig, Proteintech, China), Myc (1:1000, Cat No. 60003-2-Ig, Proteintech, China).

Techniques: Expressing, Quantitative RT-PCR, Two Tailed Test