Journal: Frontiers in Immunology

Article Title: Deciphering hub genes and immune landscapes related to neutrophil extracellular traps in rheumatoid arthritis: insights from integrated bioinformatics analyses and experiments

doi: 10.3389/fimmu.2024.1521634

Figure Lengend Snippet: Validation of pivotal gene expression in the mouse model. (A) H&E-stained images of synovial tissues of a mouse model. Expression of CRYBG1 (B) , RRM2 (C) , MMP1 (D) and SLC19A2 (E) proteins in synovial tissues of mouse knee joints was detected by IHC (standard bar = 50 μm). Data are expressed as mean ± standard deviation. *** P < 0.001, **** P < 0.0001.

Article Snippet: Next, primary antibodies CRYBG1 (1:300, bs-9093R, bioss), RRM2 (1:300, bs-7133R, bioss), MMP1 (1:100, ab52631, abcam), and SLC19A2 (1:300, bs-10738R, bioss) were added dropwise to the sections and placed in the incubator at 37°C for 60 min.

Techniques: Expressing, Staining, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Deciphering hub genes and immune landscapes related to neutrophil extracellular traps in rheumatoid arthritis: insights from integrated bioinformatics analyses and experiments

doi: 10.3389/fimmu.2024.1521634

Figure Lengend Snippet: Validation of NETs formation and hub gene expression in a clinical cohort. (A, B) Immunofluorescence microscopy examination revealed the presence of NETs in RA, defined as MPO (green) and NE (red) (standard bar = 50 μm). RT-qPCR was applied to detect the expression levels of hub genes CRYBG1 (C) , RRM2 (D) , MMP1 (E) and SLC19A2 (F) in neutrophils. *** P < 0.001, **** P < 0.0001.

Article Snippet: Next, primary antibodies CRYBG1 (1:300, bs-9093R, bioss), RRM2 (1:300, bs-7133R, bioss), MMP1 (1:100, ab52631, abcam), and SLC19A2 (1:300, bs-10738R, bioss) were added dropwise to the sections and placed in the incubator at 37°C for 60 min.

Techniques: Expressing, Immunofluorescence, Microscopy, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Deciphering hub genes and immune landscapes related to neutrophil extracellular traps in rheumatoid arthritis: insights from integrated bioinformatics analyses and experiments

doi: 10.3389/fimmu.2024.1521634

Figure Lengend Snippet: NETs-related hub genes are clinically relevant. (A) Forest plot of logistic regression analysis of hub genes in RA prediction. (B) The clinical RA prediction nomogram model is based on 4 NETs-related hub genes. (C) ROC curves of the nomogram model and the characteristic variables within the model. Calibration curves (D) , DCA (E) and CIC (F) were predicted by the nomogram model. (G-P) Correlation analysis between CRYBG1, RRM2, MMP1, and SLC19A2 and laboratory markers in RA patients. * P < 0.05, ** P < 0.01.

Article Snippet: Next, primary antibodies CRYBG1 (1:300, bs-9093R, bioss), RRM2 (1:300, bs-7133R, bioss), MMP1 (1:100, ab52631, abcam), and SLC19A2 (1:300, bs-10738R, bioss) were added dropwise to the sections and placed in the incubator at 37°C for 60 min.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Integration of scRNA-seq and ST-seq identifies hyperproliferative RRM2+ cells features and therapeutic targets in gastric cancer

doi: 10.1186/s12967-025-06847-y

Figure Lengend Snippet: Single-cell transcriptome map of cell types in GC and normal gastric specimens. ( A ) Schematic of study design. ( B ) UMAP plot of 158,622 gastric cells, colored by different groups and ( C ) different clusters. ( D ) UMAP shows 15 identified cell types in GC and normal gastric tissue, colored by cell annotation. ( E ) Heatmap displaying the top five discriminative genes for 15 different cell types. Red indicates the high expression. The box on the right presents the GO enrichment analysis results for the top-five genes. ( F ) The proportion of 15 cell types in two groups. ( G ) Expression level of RRM2 in GC and normal gastric tissues

Article Snippet: Antibody against Alpha-Tubulin (11224-1-AP), and RRM2 (11661-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Integration of scRNA-seq and ST-seq identifies hyperproliferative RRM2+ cells features and therapeutic targets in gastric cancer

doi: 10.1186/s12967-025-06847-y

Figure Lengend Snippet: Characteristics of GC epithelial cells during GC progression by trajectory analysis. ( A ) and ( B ) UMAP plots of epithelial and RRM2 + cells, colored by cell types (A) and in the two groups (B). ( C ) Trajectory analysis suggests a potential transformation pathway from epithelial cells to RRM2 + cells. ( D ) Representative gene expression in GC epithelial cells during GC progression. The color intensity reflects the normalized expression levels of genes. ( E ) Heatmap showing the change trend of MKI67, RRM2, PCNA, and TOP2A across pseudotime

Article Snippet: Antibody against Alpha-Tubulin (11224-1-AP), and RRM2 (11661-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Transformation Assay, Gene Expression, Expressing

Journal: Journal of Translational Medicine

Article Title: Integration of scRNA-seq and ST-seq identifies hyperproliferative RRM2+ cells features and therapeutic targets in gastric cancer

doi: 10.1186/s12967-025-06847-y

Figure Lengend Snippet: Analysis results combining scRNA-seq and ST. ( A ) Distribution of 15 cell types on ST slices from GC tissues ( N = 6), represented by pie charts showing the proportion of different cell types in each spot. ( B ) Distribution of RRM2 and classical proliferation markers in GC tissue sections ( N = 6), with the color getting yellow indicating a higher expression of the genes in that spot

Article Snippet: Antibody against Alpha-Tubulin (11224-1-AP), and RRM2 (11661-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Integration of scRNA-seq and ST-seq identifies hyperproliferative RRM2+ cells features and therapeutic targets in gastric cancer

doi: 10.1186/s12967-025-06847-y

Figure Lengend Snippet: Osalmid inhibits RRM2 to trigger ferroptosis and inhibits GPX4, SLC7A11, and FTH1 expression in gastric cancer. ( A ) The volcano plot showed the upregulated and downregulated genes in MKN45 cells treated with osalmid, as compared to NC (untreated with osalmid). ( B ) GO analysis demonstrated that the osalmid treatment group was engaged in the rRNA processing, ribosome biogenesis, ribonucleoprotein complex biogenesis, and so on. ( C ) KEGG analysis revealed that ferroptosis and the p53 signaling pathway were significantly enriched in the osalmid group. ( D ) Western blot analysis was performed to detect the expression differences of RRM2, FTH1, SLC7A11, GPX4, p53, and p-p53 in MKN45 and MKN7 cells with or without osalmid treatment. ( E ) CCK-8 assay showed the proliferation curve of the MKN7 and MKN45 cells after osalmid treatment or not. ( F ) Cell proliferation in the MKN7 and MKN45 were assessed by colony formation assay after osalmid treatment or not. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001

Article Snippet: Antibody against Alpha-Tubulin (11224-1-AP), and RRM2 (11661-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Western Blot, CCK-8 Assay, Colony Assay

Journal: Journal of Translational Medicine

Article Title: Integration of scRNA-seq and ST-seq identifies hyperproliferative RRM2+ cells features and therapeutic targets in gastric cancer

doi: 10.1186/s12967-025-06847-y

Figure Lengend Snippet: The effects of knocking down RRM2 on cell proliferation, colony, and ferroptosis. ( A ) Western blot of RRM2, GPX4, SLC7A11, FTH1, p53, p-p53, and ATM expression levels in RRM2 knockdown and their control cells. ( B ) The CCK-8 assay showed the proliferation curve of the MKN7 and MKN45 cells after transfection with RRM2-targeting siRNA or not. ( C ) Cell proliferation was assessed by colony formation assay in RRM2 knockdown in GC cell lines. ( D ) Comparison of GSH/GSSG ratios between siRRM2 and their control group in MKN45 and MKN7 cells. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001

Article Snippet: Antibody against Alpha-Tubulin (11224-1-AP), and RRM2 (11661-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Western Blot, Expressing, Knockdown, Control, CCK-8 Assay, Transfection, Colony Assay, Comparison

Journal: Journal of Translational Medicine

Article Title: Integration of scRNA-seq and ST-seq identifies hyperproliferative RRM2+ cells features and therapeutic targets in gastric cancer

doi: 10.1186/s12967-025-06847-y

Figure Lengend Snippet: Silencing p53 alleviates ferroptosis triggered by RRM2 loss in gastric cancer cells. ( A , B ) Knockdown of p53 in the context of RRM2 deficiency reversed the RRM2 knockdown-induced suppression of SLC7A11, FTH1, and GPX4 expression in MKN7 and MKN45. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001

Article Snippet: Antibody against Alpha-Tubulin (11224-1-AP), and RRM2 (11661-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Knockdown, Expressing

Journal: Journal of Translational Medicine

Article Title: Integration of scRNA-seq and ST-seq identifies hyperproliferative RRM2+ cells features and therapeutic targets in gastric cancer

doi: 10.1186/s12967-025-06847-y

Figure Lengend Snippet: Osalmid-induced ferroptosis in GC in vivo. ( A ) The flow charts design for in vivo experiments. ( B ) Growth curves of subcutaneous tumors in three groups. ( C ) Overview of subcutaneous tumor in three groups after 12 days. ( D ) Tumor weight and volume were assessed in three groups. ( E ) WB quantified the expression of RRM2, GPX4, SLC7A11, FTH1, p53, and p-p53 in tumor tissues in the different groups. ( F ) Comparison of GSH/GSSG ratios between shRRM2 or Osalmid and their control groups. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001

Article Snippet: Antibody against Alpha-Tubulin (11224-1-AP), and RRM2 (11661-1-AP) were purchased from Proteintech (Wuhan, China).

Techniques: In Vivo, Expressing, Comparison, Control

Journal: bioRxiv

Article Title: Translational evidence for RRM2 as a prognostic biomarker and therapeutic target in Ewing sarcoma

doi: 10.1101/2021.03.04.433896

Figure Lengend Snippet: a) Schematic description of the filtering process for identification of therapeutically relevant target candidates. b) Analysis of RRM2 mRNA expression levels in 50 EwS primary tumors compared to 929 normal tissues samples from 71 tissue types. Data are shown as log2 fold increase normalized to expression values of normal tissues. The dotted line indicates the cut-off value of 2 for candidate selection. c) Analysis of overall survival time of 166 EwS patients stratified for candidate gene expression. P -values (–log10) were determined in Kaplan-Meier analyses using a Mantel-Haenszel test (Bonferroni-adjusted for multiple testing). The dotted line indicates a significance value of 1.3. d) Kaplan-Meier survival analysis of 166 EwS patients stratified by the 78th percentile RRM2 expression. P -value determined by log-rank test.

Article Snippet: Slides were incubated with a polyclonal anti-RRM2 primary antibody (rabbit, 1:500, atlas antibodies, HPA056994) for 60 min at room temperature followed by incubation with a monoclonal secondary horseradish peroxidase (HRP)-coupled horse-anti-rabbit antibody with AEC+ as chromogen, counterstained by hematoxylin Gill’s formula (H-3401, Vector Laboratories, Germany).

Techniques: Expressing, Selection, Gene Expression

Journal: bioRxiv

Article Title: Translational evidence for RRM2 as a prognostic biomarker and therapeutic target in Ewing sarcoma

doi: 10.1101/2021.03.04.433896

Figure Lengend Snippet: a) Left: Heat map for gene expression which is positively or negatively correlated with RRM2 expression in 166 EwS. Right: Gene ontology (GO) enrichment analysis of RRM2 and its co-expressed genes derived from gene expression data sets of 166 EwS tumors. Pearson correlation coefficients between RRM2 and other genes were determined, of which those with |r Pearson | > 0.5 were further analyzed by GO enrichment analysis. b) Representative images of immunohistochemical RRM2 staining. Scale bar = 50 µm. c) Kaplan-Meier survival analysis of 122 EwS patients stratified by RRM2 protein expression (low IRS≤2, high IRS >2). P -values were determined by log-rank test. d) WGCNA of downregulated genes upon RRM2 silencing in A-673 and ES-7 cells harboring Dox-inducible shRRM2 constructs. NES, normalized enrichment score. e) Analysis of tumor growth of EwS cell lines A-673 and TC-71 harboring Dox-inducible shRRM2 constructs or non-targeting shRNA (shControl) xenografted in NSG mice. Once tumors were palpable, animals were randomized in Dox (+) or Dox (–) group. Tumor growth on time course and f) Tumor weight at the experimental endpoint. Arrows indicate treatment start. Values are normalized to shControl. Horizontal bars represent means and whiskers SEM. FC, fold change. P -values were calculated at the experimental endpoint with two-sided (tumor growth) or one-sided (tumor weight) Mann-Whitney test. g) Representative micrographs of xenografts immunohistochemically stained for RRM2, cleaved caspase-3 (CC3) or γH2A.X (scale bar=250 µm, 50 µm, 250 µm, respectively). h) quantification of positive cells for cleaved caspase-3 (CC3) (left) and γH2A.X (right). Values were normalized to shControl. Horizontal bars represent means and whiskers SEM. FC, fold change. P -values were calculated at the experimental endpoint using a two-sided Mann-Whitney test.

Article Snippet: Slides were incubated with a polyclonal anti-RRM2 primary antibody (rabbit, 1:500, atlas antibodies, HPA056994) for 60 min at room temperature followed by incubation with a monoclonal secondary horseradish peroxidase (HRP)-coupled horse-anti-rabbit antibody with AEC+ as chromogen, counterstained by hematoxylin Gill’s formula (H-3401, Vector Laboratories, Germany).

Techniques: Gene Expression, Expressing, Derivative Assay, Immunohistochemical staining, Staining, Construct, shRNA, MANN-WHITNEY

Journal: bioRxiv

Article Title: Translational evidence for RRM2 as a prognostic biomarker and therapeutic target in Ewing sarcoma

doi: 10.1101/2021.03.04.433896

Figure Lengend Snippet: a) Analysis of proliferation assays upon shRNA-mediated RRM2 silencing in EwS cell lines. Upper: Cell proliferation over 120h upon RRM2 silencing in A-673. Viable cells upon RRM2 silencing (middle) and dead cells (lower) in EwS cell lines (A-673, ES7, TC-71) harboring Dox-inducible shRRM2 constructs or non-targeting shRNA (shControl). Values are normalized to shControl. Horizontal bars represent means and whiskers SEM. FC, fold change. Two-sided Mann-Whitney test. b) Analysis of clonogenic growth upon shRNA-mediated RRM2 silencing in EwS cell lines (A-673, ES7, TC-71) harboring Dox-inducible shRRM2 constructs or non-targeting shRNA (shControl). Horizontal bars represent means and whiskers SEM. Two-sided Mann-Whitney test at the experimental endpoint.

Article Snippet: Slides were incubated with a polyclonal anti-RRM2 primary antibody (rabbit, 1:500, atlas antibodies, HPA056994) for 60 min at room temperature followed by incubation with a monoclonal secondary horseradish peroxidase (HRP)-coupled horse-anti-rabbit antibody with AEC+ as chromogen, counterstained by hematoxylin Gill’s formula (H-3401, Vector Laboratories, Germany).

Techniques: shRNA, Construct, MANN-WHITNEY

Journal: bioRxiv

Article Title: Translational evidence for RRM2 as a prognostic biomarker and therapeutic target in Ewing sarcoma

doi: 10.1101/2021.03.04.433896

Figure Lengend Snippet: a) Integrative Gene Ontology (GO) enrichment analysis of gene expression microarray data generated in A-673 and ES7 cells after RRM2 silencing or pharmacological RRM2 inhibition by triapine (corresponding IC50 of 0.44 µM or 0.65 µM, respectively). b) Correlation of gene expression between RRM2 and CHEK1 or WEE1 in 166 EwS. Each dot represents an individual expression value. Solid red lines indicate a trend line created by a simple linear regression. P -values were calculated by a two-tailed t-test. c) Drug interaction and combination efficiency analysis between triapine and CHEK1 inhibitor (CCT245737) or WEE1 inhibitor (MK-1775) in 4 EwS cell lines (A-673, ES7, EW-7, TC-71) assessed by combination index. CI value < 1 indicative of synergistic, CI = 1 additive, and CI > 1 antagonistic d) Drug interaction and combination efficiency estimation between triapine and CHEK1 inhibitor (CCT245737) or WEE1 inhibitor (MK-1775) in A-673 EwS cell line assessed by SynergyFinder 2.0. ZIP synergy score > 10, likely to be synergistic; between -10 and 10, likely to be additive; < –10, likely to be antagonistic.

Article Snippet: Slides were incubated with a polyclonal anti-RRM2 primary antibody (rabbit, 1:500, atlas antibodies, HPA056994) for 60 min at room temperature followed by incubation with a monoclonal secondary horseradish peroxidase (HRP)-coupled horse-anti-rabbit antibody with AEC+ as chromogen, counterstained by hematoxylin Gill’s formula (H-3401, Vector Laboratories, Germany).

Techniques: Gene Expression, Microarray, Generated, Inhibition, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Translational evidence for RRM2 as a prognostic biomarker and therapeutic target in Ewing sarcoma

doi: 10.1101/2021.03.04.433896

Figure Lengend Snippet: a) Distribution analysis of RRM2 mRNA expression in 166 EwS patients. Each dot represents individual RRM2 expression. b) Correlation analysis of CNVs at the RRM2 locus with RRM2 mRNA expression levels in primary EwS tumors (n=32). The solid line indicates a trend line estimated by a simple linear regression model. b) Correlation analysis of promoter methylation on five CpG sites with RRM2 expression levels in primary EwS tumors (n=40). The solid lines indicate trend lines estimated by a simple linear regression model.

Article Snippet: Slides were incubated with a polyclonal anti-RRM2 primary antibody (rabbit, 1:500, atlas antibodies, HPA056994) for 60 min at room temperature followed by incubation with a monoclonal secondary horseradish peroxidase (HRP)-coupled horse-anti-rabbit antibody with AEC+ as chromogen, counterstained by hematoxylin Gill’s formula (H-3401, Vector Laboratories, Germany).

Techniques: Expressing, Methylation

Journal: Heliyon

Article Title: Tanshinone IIA attenuates fluoride-induced spinal cord injury by inhibiting ferroptosis and inflammation

doi: 10.1016/j.heliyon.2024.e40549

Figure Lengend Snippet: Primers used in qRT-PCR study.

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against CXCL2 (26791-1-AP, Proteintech), PCK2 (67676-1-lg, abcam), RRM2 (DF7248, Affinity), SLC7A11 (BM5318, BOSTER), or β-actin (BA2305, BOSTER).

Techniques:

Journal: Heliyon

Article Title: Tanshinone IIA attenuates fluoride-induced spinal cord injury by inhibiting ferroptosis and inflammation

doi: 10.1016/j.heliyon.2024.e40549

Figure Lengend Snippet: Tanshinone IIA improved spinal cord injury by inhibiting ferroptosis (A) Photomicrographs of spinal cord sections from Tanshinone IIA control, fluoride-exposed rats, and Tanshinone IIA + high fluoride group visualized by transmission electron microscopy (5000X, white box). Fibrous myelin integrity (indicated by red arrows) is maintained in both Tanshinone IIA control and Tanshinone IIA-treated groups, while in the fluorosis group, it appears loosely arranged with an increased gap and disordered structure. (B) Scatter plot exhibited the expression of 15 ferroptosis-related genes in Tan IIA control, high fluoride group, and Tan IIA + high fluoride group. (C–D) Western blots showed that Tan IIA treatment restored the reduced protein expression of SLC7A11 and RRM2 observed in the NaF group. Additionally, Tan IIA treatment decreased the elevated protein levels of CXCL2 and PCK2 in the NaF group. Uncropped and unadjusted original images of the Western blots are provided in . Data are represented as the mean ± SEM (n = 6). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001.

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against CXCL2 (26791-1-AP, Proteintech), PCK2 (67676-1-lg, abcam), RRM2 (DF7248, Affinity), SLC7A11 (BM5318, BOSTER), or β-actin (BA2305, BOSTER).

Techniques: Control, Transmission Assay, Electron Microscopy, Expressing, Western Blot

Journal: Biomolecules

Article Title: RRM2 Alleviates Doxorubicin-Induced Cardiotoxicity through the AKT/mTOR Signaling Pathway

doi: 10.3390/biom12020299

Figure Lengend Snippet: RRM2 expression was decreased in vitro and in vivo after DOX treatment. ( A ) Compared with the CTR group, the mRNA expression of RRM2 in primary cardiomyocytes was decreased after DOX treatment ( n = 6). ( B ) Compared with the CTR group, the mRNA expression of RRM2 in hearts was lower after DOX treatment ( n = 6). ( C , D ) RRM2 protein levels in primary cardiomyocytes treated with DOX ( n = 4). ( E , F ) RRM2 protein levels in hearts 5 days after DOX treatment ( n = 4). ** indicates p < 0.01. *** indicates p < 0.001.

Article Snippet: The antibodies we used were as follows: RRM2 (ABclonal, A5255), Bcl-2 (Proteintech, 12789-1-AP), Phospho-mTOR (CST, 5536T), cleaved-Caspase3 (CST, 9661S), Beclin 1 (ABclonal, A7353), mTOR (CST, 2983T), AKT (Proteintech, 10176-2-AP), Phospho-Akt (Abclonal, AP1259), LC3B (Abclonal, A19665), GAPDH (Abcam, ab181602).

Techniques: Expressing, In Vitro, In Vivo

Journal: Biomolecules

Article Title: RRM2 Alleviates Doxorubicin-Induced Cardiotoxicity through the AKT/mTOR Signaling Pathway

doi: 10.3390/biom12020299

Figure Lengend Snippet: RRM2 overexpression protected against apoptosis and autophagy in cardiomyocytes. ( A – E ) The protein expressions of Bcl-2, C-Caspase3, Beclin1, and LC3B in cardiomyocytes after DOX treatment ( n = 4). ( F , G ) Western blot showing RRM2 level in cardiomyocytes after virus transfection ( n = 4). ( H , I ) The protein expression of autophagy and apoptosis ( n = 4). ** indicates p < 0.01. *** indicates p < 0.001.

Article Snippet: The antibodies we used were as follows: RRM2 (ABclonal, A5255), Bcl-2 (Proteintech, 12789-1-AP), Phospho-mTOR (CST, 5536T), cleaved-Caspase3 (CST, 9661S), Beclin 1 (ABclonal, A7353), mTOR (CST, 2983T), AKT (Proteintech, 10176-2-AP), Phospho-Akt (Abclonal, AP1259), LC3B (Abclonal, A19665), GAPDH (Abcam, ab181602).

Techniques: Over Expression, Western Blot, Virus, Transfection, Expressing

Journal: Biomolecules

Article Title: RRM2 Alleviates Doxorubicin-Induced Cardiotoxicity through the AKT/mTOR Signaling Pathway

doi: 10.3390/biom12020299

Figure Lengend Snippet: RRM2 knockdown facilitated DOX-induced injury in vitro. ( A ) The mRNA expression of RRM2 after si-RNA transfection ( n = 6). ( B , C ) The protein expression of RRM2 after si-2 transfection ( n = 4). ( D , E ) The protein expression of autophagy and apoptosis after si-2 and DOX treatment ( n = 4). ( F ) Cell viability after Ad-RRM2, si-2, and DIDOX treatment. ( n = 5). * indicates p < 0.05. ** indicates p < 0.01. *** indicates p < 0.001. **** indicates p < 0.0001.

Article Snippet: The antibodies we used were as follows: RRM2 (ABclonal, A5255), Bcl-2 (Proteintech, 12789-1-AP), Phospho-mTOR (CST, 5536T), cleaved-Caspase3 (CST, 9661S), Beclin 1 (ABclonal, A7353), mTOR (CST, 2983T), AKT (Proteintech, 10176-2-AP), Phospho-Akt (Abclonal, AP1259), LC3B (Abclonal, A19665), GAPDH (Abcam, ab181602).

Techniques: Knockdown, In Vitro, Expressing, Transfection

Journal: Biomolecules

Article Title: RRM2 Alleviates Doxorubicin-Induced Cardiotoxicity through the AKT/mTOR Signaling Pathway

doi: 10.3390/biom12020299

Figure Lengend Snippet: RRM2 overexpression alleviated DOX-induced cardiotoxicity in vivo. ( A , B ) Immunofluorescence data revealed that Ad-GFP and Ad-RRM2 were successfully transferred into hearts ( n = 4). ( C , D ) The protein levels of autophagy and apoptosis in mice after DOX treatment ( n = 4). ( E ) H&E staining in hearts compared with DOX and adenovirus treatments. ** indicates p < 0.01. *** indicates p < 0.001.

Article Snippet: The antibodies we used were as follows: RRM2 (ABclonal, A5255), Bcl-2 (Proteintech, 12789-1-AP), Phospho-mTOR (CST, 5536T), cleaved-Caspase3 (CST, 9661S), Beclin 1 (ABclonal, A7353), mTOR (CST, 2983T), AKT (Proteintech, 10176-2-AP), Phospho-Akt (Abclonal, AP1259), LC3B (Abclonal, A19665), GAPDH (Abcam, ab181602).

Techniques: Over Expression, In Vivo, Immunofluorescence, Staining

Journal: Biomolecules

Article Title: RRM2 Alleviates Doxorubicin-Induced Cardiotoxicity through the AKT/mTOR Signaling Pathway

doi: 10.3390/biom12020299

Figure Lengend Snippet: Blocking the overexpression of RRM2 reversed its protective effect. ( A , B ) The protein levels of Bcl-2, C-Caspase3, Beclin1, and LC3B in cardiomyocytes after DIDOX and adenovirus treatments ( n = 4). ( C ) TUNEL staining in each group ( n = 4). ** indicates p < 0.01. *** indicates p < 0.001.

Article Snippet: The antibodies we used were as follows: RRM2 (ABclonal, A5255), Bcl-2 (Proteintech, 12789-1-AP), Phospho-mTOR (CST, 5536T), cleaved-Caspase3 (CST, 9661S), Beclin 1 (ABclonal, A7353), mTOR (CST, 2983T), AKT (Proteintech, 10176-2-AP), Phospho-Akt (Abclonal, AP1259), LC3B (Abclonal, A19665), GAPDH (Abcam, ab181602).

Techniques: Blocking Assay, Over Expression, TUNEL Assay, Staining

Journal: Biomolecules

Article Title: RRM2 Alleviates Doxorubicin-Induced Cardiotoxicity through the AKT/mTOR Signaling Pathway

doi: 10.3390/biom12020299

Figure Lengend Snippet: The effects of RRM2 on cardiotoxicity induced by DOX.

Article Snippet: The antibodies we used were as follows: RRM2 (ABclonal, A5255), Bcl-2 (Proteintech, 12789-1-AP), Phospho-mTOR (CST, 5536T), cleaved-Caspase3 (CST, 9661S), Beclin 1 (ABclonal, A7353), mTOR (CST, 2983T), AKT (Proteintech, 10176-2-AP), Phospho-Akt (Abclonal, AP1259), LC3B (Abclonal, A19665), GAPDH (Abcam, ab181602).

Techniques:

Journal: iScience

Article Title: Regorafenib promotes antitumor progression in melanoma by reducing RRM2

doi: 10.1016/j.isci.2024.110993

Figure Lengend Snippet: RRM2 was the downstream target of regorafenib (A and B) PCA suggested a clear distinction between regorafenib treatment and control samples. (C) 1502 upregulated and 780 downregulated DEGs were identified in regorafenib treatment samples by RNA-seq. (D and E) Downregulated DEGs were used for enrichment analysis. (F) 25 ODEGs were identified through combining analysis of our RNA-seq_down genes and GSE7553 _up genes. (G) The levels of RRM2 protein were suppressed when melanoma cells were treated with different concentrations of regorafenib. (H) The predicted binding mode of regorafenib to RRM2 by docking analysis. Reg: regorafenib. ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Data are represented as mean ± SEM. Similar results were obtained in two additional independent experiments.

Article Snippet: The primary antibodies were as follows: anti-PARP/cleaved-PARP (1:1000, 13371-1-AP, Proteintech, USA), Bax (1:5000, 50599-2-Ig, Proteintech, USA), Bcl-2 (1:5000, 12789-1-AP, Proteintech, USA), anti-phospho-p44/42 MAPK (Erk1/2) (1:1000, 4370, Cell Signaling Technology, USA), anti-p44/42 MAPK (Erk1/2) (1:1000, 4695, Cell Signaling Technology, USA), RRM2 (1:1000, A5504, Bimake, USA), E2F3 (1:1000, DF12390, Affinity Biosciences, USA) and β-actin (1:3000, 20536-1-AP, Proteintech, USA).

Techniques: Control, RNA Sequencing Assay, Binding Assay

Journal: iScience

Article Title: Regorafenib promotes antitumor progression in melanoma by reducing RRM2

doi: 10.1016/j.isci.2024.110993

Figure Lengend Snippet: Depletion of RRM2 alleviated the malignancy of melanoma cells (A) The depletion efficiency of RRM2 was confirmed by western blot analysis. (B–E) Depletion of RRM2 significantly inhibited the proliferation of Sk-Mel-2/28 cells by CCK8 (B) and EDU staining (C–E). (F–I) RRM2 depletion reduced the number of invaded cells (F–H) and retarded the wound closure of melanoma cells (I). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Data are represented as mean ± SEM. Similar results were obtained in two additional independent experiments. (C + F) Scale bar: 100 μm.

Article Snippet: The primary antibodies were as follows: anti-PARP/cleaved-PARP (1:1000, 13371-1-AP, Proteintech, USA), Bax (1:5000, 50599-2-Ig, Proteintech, USA), Bcl-2 (1:5000, 12789-1-AP, Proteintech, USA), anti-phospho-p44/42 MAPK (Erk1/2) (1:1000, 4370, Cell Signaling Technology, USA), anti-p44/42 MAPK (Erk1/2) (1:1000, 4695, Cell Signaling Technology, USA), RRM2 (1:1000, A5504, Bimake, USA), E2F3 (1:1000, DF12390, Affinity Biosciences, USA) and β-actin (1:3000, 20536-1-AP, Proteintech, USA).

Techniques: Western Blot, Staining

Journal: iScience

Article Title: Regorafenib promotes antitumor progression in melanoma by reducing RRM2

doi: 10.1016/j.isci.2024.110993

Figure Lengend Snippet: RRM2 overexpression partially reversed regorafenib-induced growth inhibition of melanoma cells (A) RRM2 overexpression partially reversed regorafenib-induced cytotoxicity of melanoma cells by CCK8 cytotoxicity test. (B and C) RRM2 overexpression partially reduced regorafenib-induced apoptosis of melanoma cells by FCM. (D and E) RRM2 overexpression partially reverses the suppression of invasion and migration induced by regorafenib in melanoma cells by transell assays. Scale bar: 100 μm. Reg: regorafenib. ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Data are represented as mean ± SEM. Similar results were obtained in two additional independent experiments.

Article Snippet: The primary antibodies were as follows: anti-PARP/cleaved-PARP (1:1000, 13371-1-AP, Proteintech, USA), Bax (1:5000, 50599-2-Ig, Proteintech, USA), Bcl-2 (1:5000, 12789-1-AP, Proteintech, USA), anti-phospho-p44/42 MAPK (Erk1/2) (1:1000, 4370, Cell Signaling Technology, USA), anti-p44/42 MAPK (Erk1/2) (1:1000, 4695, Cell Signaling Technology, USA), RRM2 (1:1000, A5504, Bimake, USA), E2F3 (1:1000, DF12390, Affinity Biosciences, USA) and β-actin (1:3000, 20536-1-AP, Proteintech, USA).

Techniques: Over Expression, Inhibition, Migration

Journal: iScience

Article Title: Regorafenib promotes antitumor progression in melanoma by reducing RRM2

doi: 10.1016/j.isci.2024.110993

Figure Lengend Snippet: ERK/E2F3 signaling participated the inhibition of regorafenib on melanoma cells (A) The expression of p-ERK and E2F3 were inhibited when melanoma cells were treated with regorafenib by western blot. (B) When ERK signaling was blocked, the level of p-ERK, E2F3, and RRM2 was reduced. (C) When E2F3 expression was depleted, RRM2 expression was also depleted. Reg: regorafenib. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Data are represented as mean ± SEM. Similar results were obtained in two additional independent experiments.

Article Snippet: The primary antibodies were as follows: anti-PARP/cleaved-PARP (1:1000, 13371-1-AP, Proteintech, USA), Bax (1:5000, 50599-2-Ig, Proteintech, USA), Bcl-2 (1:5000, 12789-1-AP, Proteintech, USA), anti-phospho-p44/42 MAPK (Erk1/2) (1:1000, 4370, Cell Signaling Technology, USA), anti-p44/42 MAPK (Erk1/2) (1:1000, 4695, Cell Signaling Technology, USA), RRM2 (1:1000, A5504, Bimake, USA), E2F3 (1:1000, DF12390, Affinity Biosciences, USA) and β-actin (1:3000, 20536-1-AP, Proteintech, USA).

Techniques: Inhibition, Expressing, Western Blot

Journal: iScience

Article Title: Regorafenib promotes antitumor progression in melanoma by reducing RRM2

doi: 10.1016/j.isci.2024.110993

Figure Lengend Snippet: Regorafenib reduced tumorigenesis in vivo (A–C) Regorafenib inhibited the growth of xenograft tumors, including tumor weight (B) and volume (C). (D) Regorafenib treatment inhibited the levels of p-ERK and RRM2, while RRM2 overexpression partially relieved the inhibition of p-ERK and RRM2 expression. (E) There were no obvious differences in the weight of these vital organs between control and regorafenib-treatment group. ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Data are represented as mean ± SEM. Similar results were obtained in two additional independent experiments.

Article Snippet: The primary antibodies were as follows: anti-PARP/cleaved-PARP (1:1000, 13371-1-AP, Proteintech, USA), Bax (1:5000, 50599-2-Ig, Proteintech, USA), Bcl-2 (1:5000, 12789-1-AP, Proteintech, USA), anti-phospho-p44/42 MAPK (Erk1/2) (1:1000, 4370, Cell Signaling Technology, USA), anti-p44/42 MAPK (Erk1/2) (1:1000, 4695, Cell Signaling Technology, USA), RRM2 (1:1000, A5504, Bimake, USA), E2F3 (1:1000, DF12390, Affinity Biosciences, USA) and β-actin (1:3000, 20536-1-AP, Proteintech, USA).

Techniques: In Vivo, Over Expression, Inhibition, Expressing, Control

Journal: iScience

Article Title: Regorafenib promotes antitumor progression in melanoma by reducing RRM2

doi: 10.1016/j.isci.2024.110993

Figure Lengend Snippet:

Article Snippet: The primary antibodies were as follows: anti-PARP/cleaved-PARP (1:1000, 13371-1-AP, Proteintech, USA), Bax (1:5000, 50599-2-Ig, Proteintech, USA), Bcl-2 (1:5000, 12789-1-AP, Proteintech, USA), anti-phospho-p44/42 MAPK (Erk1/2) (1:1000, 4370, Cell Signaling Technology, USA), anti-p44/42 MAPK (Erk1/2) (1:1000, 4695, Cell Signaling Technology, USA), RRM2 (1:1000, A5504, Bimake, USA), E2F3 (1:1000, DF12390, Affinity Biosciences, USA) and β-actin (1:3000, 20536-1-AP, Proteintech, USA).

Techniques: Imaging, Cell Cycle Assay, SYBR Green Assay, Software