Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Analysis and validation of signature genes. (A) Violin plot of characteristic gene expression in normal and OA samples. (B) Line plot of signature gene expression in normal and OA samples. (C) ROC curves to validate the diagnostic accuracy of signature genes. (D) Expression of ERBB2 in the GSE48556 dataset.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Biomarker Discovery, Gene Expression, Diagnostic Assay, Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Analysis of ERBB2 expression. (A) Heat maps of the top 20 DEGs in high and low ERBB2 expression groups. (B) Volcano map of DEGs in ERBB2 high and low expression groups. (D) Heat map of the correlation between ERBB2 and DEGs, with positive correlation in red and negative correlation in blue.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Immune-related function analysis in samples of high and low ERBB2 expression groups (* p < 0.05, ** p < 0.01).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: GSEA and GSVA analysis of ERBB2. (A–B) GSEA analysis of ERBB2 in high expression group. (C–D) GSEA analysis of ERBB2 in low expression group. (E) E GSVA analysis of biological processes, cellular components, and molecular functions of ERBB2. (F) Pathway-related GSVA analysis of ERBB2.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: ceRNA Regulatory network and potential drug action network. (A) ceRNA network regulation diagram of lncRNA-miRNA-ERBB2. (B) Potential drug-signature gene network diagram of OA, red for ERBB2, blue for drug.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques:

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: ERBB2 Transcription levels and protein expression levels between normal and OA samples. (A) Expression level of ERBB2 in qRT-PCR. (B) Protein expression level of ERBB2. (C) Immunohistochemical results of normal human cartilage and OA cartilage from patients (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Tissue staining and immunohistochemical analysis. (A) Hematoxylin-eosin staining of normal and OA rat knee joints. (B) so/FG staining of normal and OA rat knee joints. (C) OARSI score. (D-E) Protein expression level and quantitative analysis of ERBB2 in knee cartilage by IHC (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Staining, Immunohistochemical staining, Expressing, Paraffin-embedded Immunohistochemistry

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Inhibition of Tumor Growth and Immunomodulatory Effects of Flavonoids and Scutebarbatines of Scutellaria barbata D. Don in Lewis-Bearing C57BL/6 Mice

doi: 10.1155/2015/630760

Figure Lengend Snippet: Effect of flavonoids and scutebarbatines in SB on the levels of immune factors in Lewis tumor-bearing mice. All data were expressed as means ± SD ( n = 9).

Article Snippet: Antibodies to these immunoregulatory factors including IL-10, IL-17, FOXP3, IFN- γ , IL-2, ROR γ t, TGF- β 1, IL-6, and β -actin were obtained from Wuhan BOSTER Biological Engineering Co., Ltd. (Wuhan, China).

Techniques:

Journal: eLife

Article Title: Unique integrated stress response sensors regulate cancer cell susceptibility when Hsp70 activity is compromised

doi: 10.7554/eLife.64977

Figure Lengend Snippet: ( A ) HER2 (diamonds), TNBC (circles), and luminal (squares and triangles) breast cancer lines were seeded into 96-well plates and treated with increasing doses of MAL3-101 for 72 hr. Viability is expressed as the average of three or more independent experiments, ± SEM. ( B ) MAL3-101-sensitive (MCF7 and MDA MB 231, denoted in blue) and resistant (MDA MB 453 and MDA MB 361, denoted in black) cells were treated with 12 µM MAL3-101 for the indicated times, and lysates were prepared and immunoblotted for cleaved caspase-3, caspase-7, and caspase-8. β-actin serves as a loading control. ( C ) The corresponding fold-increase of the indicated apoptotic markers relative to the DMSO control are plotted, ± SEM (n≥3 for cleaved caspase-3, n=3 for cleaved caspase-7, and n≥4 for cleaved caspase-8). Black asterisks correspond to statistical significance between MDA MB 231 cells (closed circle) and MDA MB 453 and MDA MB 361 (open circle and triangle, respectively), and the red asterisk represents statistical significance between MCF7 (closed triangle) and MDA MB 453 and MDA MB 361 (open circle and triangle) cells; * denotes p<0.05, ** denotes p<0.005. Figure 1—source data 1. Source data for cell viability assay and apoptotic marker accumulation in .

Article Snippet: For the remaining proteins aliquots from the same lysates were instead heated to 37°C for 30 min prior to SDS-PAGE using 4–20% Tris-Glycine gradient gels (XP04202Box, Thermo Fisher Scientific), and after transfer on nitrocellulose membranes (#84–874, Prometheus Laboratories Inc, California), the blots were incubated with anti-cleaved Caspase-8 (18C8, #9496; at 1:1000), anti-BiP (C50B12, #3177S; at 1:1000), anti-Hsp70 (smc-113, StressMarq Biosciences, Victoria, British Columbia; at 1:1000), anti-Hsc/Hsp70 (#4872S; at 1:1000), anti-HER2 (29D8, #2165; at 1:2000), anti-eIF2α (#9722S; at 1:2000), anti-phospho-eIF2α (#9721L; at 1:500), anti-PERK (D11A8, #5683S; at 1:2000), anti-PKR (D7F7, #12297S; at 1:2000), anti-Ire1α (14C10, #3294S; at 1:1000), anti-HRI (MBS2538114, MyBioSource, San Diego, CA; at 1:500) and anti-GCN2 and anti-GCN2 pT899 antibody (ab-134053 and ab-75836, Abcam, Cambtidge, UK; at 1:2000).

Techniques: Viability Assay, Marker

Journal: eLife

Article Title: Unique integrated stress response sensors regulate cancer cell susceptibility when Hsp70 activity is compromised

doi: 10.7554/eLife.64977

Figure Lengend Snippet: Breast cancer cells exhibit a range of sensitivities to MAL3-101, a specific Hsp70 inhibitor. The indicated breast cancer lines were seeded into 96-well plates and treated with increasing doses of the indicated compounds for 72 hr. Viability was measured using the CellTiter-Glo assay. IC 50 values were generate using a sigmoidal nonlinear regression with SigmaPlot 11.0. ND stands for an undetermined value. MAL3-101 sensitivities of Hsp70 inhibitor resistant cells are in bold.

Article Snippet: For the remaining proteins aliquots from the same lysates were instead heated to 37°C for 30 min prior to SDS-PAGE using 4–20% Tris-Glycine gradient gels (XP04202Box, Thermo Fisher Scientific), and after transfer on nitrocellulose membranes (#84–874, Prometheus Laboratories Inc, California), the blots were incubated with anti-cleaved Caspase-8 (18C8, #9496; at 1:1000), anti-BiP (C50B12, #3177S; at 1:1000), anti-Hsp70 (smc-113, StressMarq Biosciences, Victoria, British Columbia; at 1:1000), anti-Hsc/Hsp70 (#4872S; at 1:1000), anti-HER2 (29D8, #2165; at 1:2000), anti-eIF2α (#9722S; at 1:2000), anti-phospho-eIF2α (#9721L; at 1:500), anti-PERK (D11A8, #5683S; at 1:2000), anti-PKR (D7F7, #12297S; at 1:2000), anti-Ire1α (14C10, #3294S; at 1:1000), anti-HRI (MBS2538114, MyBioSource, San Diego, CA; at 1:500) and anti-GCN2 and anti-GCN2 pT899 antibody (ab-134053 and ab-75836, Abcam, Cambtidge, UK; at 1:2000).

Techniques:

Journal: eLife

Article Title: Unique integrated stress response sensors regulate cancer cell susceptibility when Hsp70 activity is compromised

doi: 10.7554/eLife.64977

Figure Lengend Snippet: The cell numbers and autophagy or proteasome inhibitor concentrations used for the cell viability assay in combination with increasing doses of MAL3-101 are shown. The concentrations of bortezomib, CQ, and bafilomycin to induce no greater than 30% of cell death in each line after 72 hr treatment are shown. ND stands for undetermined value.

Article Snippet: For the remaining proteins aliquots from the same lysates were instead heated to 37°C for 30 min prior to SDS-PAGE using 4–20% Tris-Glycine gradient gels (XP04202Box, Thermo Fisher Scientific), and after transfer on nitrocellulose membranes (#84–874, Prometheus Laboratories Inc, California), the blots were incubated with anti-cleaved Caspase-8 (18C8, #9496; at 1:1000), anti-BiP (C50B12, #3177S; at 1:1000), anti-Hsp70 (smc-113, StressMarq Biosciences, Victoria, British Columbia; at 1:1000), anti-Hsc/Hsp70 (#4872S; at 1:1000), anti-HER2 (29D8, #2165; at 1:2000), anti-eIF2α (#9722S; at 1:2000), anti-phospho-eIF2α (#9721L; at 1:500), anti-PERK (D11A8, #5683S; at 1:2000), anti-PKR (D7F7, #12297S; at 1:2000), anti-Ire1α (14C10, #3294S; at 1:1000), anti-HRI (MBS2538114, MyBioSource, San Diego, CA; at 1:500) and anti-GCN2 and anti-GCN2 pT899 antibody (ab-134053 and ab-75836, Abcam, Cambtidge, UK; at 1:2000).

Techniques: Viability Assay

Journal: eLife

Article Title: Unique integrated stress response sensors regulate cancer cell susceptibility when Hsp70 activity is compromised

doi: 10.7554/eLife.64977

Figure Lengend Snippet: Breast cancer cells exhibit a range of sensitivities to MAL3-101 in the presence of either autophagy or proteasome inhibitors. Cells were seeded into 96-well plates and treated with increasing doses of MAL3-101 in the presence or absence of subcritical doses of bortezomib (proteasome inhibitor), or CQ or bafilomycin (autophagy inhibitors) for 72 hr. Viability was measured using the CellTiter-Glo assay. IC 50 values were generate using a sigmoidal nonlinear regression with SigmaPlot 11.0. ND stands for undetermined value. MAL3-101 resistant cells are highlighted in yellow.

Article Snippet: For the remaining proteins aliquots from the same lysates were instead heated to 37°C for 30 min prior to SDS-PAGE using 4–20% Tris-Glycine gradient gels (XP04202Box, Thermo Fisher Scientific), and after transfer on nitrocellulose membranes (#84–874, Prometheus Laboratories Inc, California), the blots were incubated with anti-cleaved Caspase-8 (18C8, #9496; at 1:1000), anti-BiP (C50B12, #3177S; at 1:1000), anti-Hsp70 (smc-113, StressMarq Biosciences, Victoria, British Columbia; at 1:1000), anti-Hsc/Hsp70 (#4872S; at 1:1000), anti-HER2 (29D8, #2165; at 1:2000), anti-eIF2α (#9722S; at 1:2000), anti-phospho-eIF2α (#9721L; at 1:500), anti-PERK (D11A8, #5683S; at 1:2000), anti-PKR (D7F7, #12297S; at 1:2000), anti-Ire1α (14C10, #3294S; at 1:1000), anti-HRI (MBS2538114, MyBioSource, San Diego, CA; at 1:500) and anti-GCN2 and anti-GCN2 pT899 antibody (ab-134053 and ab-75836, Abcam, Cambtidge, UK; at 1:2000).

Techniques: