anti rfc2  (Proteintech)

Journal: Cell Reports

Article Title: Defining bottlenecks and opportunities for Lassa virus neutralization by structural profiling of vaccine-induced polyclonal antibody responses

doi: 10.1016/j.celrep.2024.114708

Figure Lengend Snippet:

Article Snippet: pFUSE-rIgG-Fc , Invivogen , Cat# pfuse-rfc1.

Techniques: Virus, Purification, Recombinant, Electron Microscopy, Lysis, Luciferase, Irradiation, Expressing, Plasmid Preparation, Software, Luminex, Mutagenesis, Nano Differential Scanning Fluorimetry, Microscopy, Clinical Proteomics

Journal: Scientific Reports

Article Title: Identification of DNA damage response-related genes as biomarkers for castration-resistant prostate cancer

doi: 10.1038/s41598-023-46651-6

Figure Lengend Snippet: Knockdown of RFC2 results in accumulation of DNA damage, increased apoptosis and growth inhibition in CRPC cells. ( A ) The expression level of RFC2 at the protein level in multiple prostate cell lines. Lysates from five prostate cell lines (RWPE, LNCaP, 22Rv1, DU145 and PC3 cells) were used for Western blot analysis with specific antibodies to replication factor C subunit2 (RFC2). β-Actin was used as a loading control. ( B ) The expression level of RFC2 at the protein level in LTAD and parental LNCaP cells. Lysates were used for Western blot analysis with specific antibodies to replication factor C subunit2 (RFC2). β-Actin was used as a loading control. ( C ) Knockdown of RFC2 suppresses cell proliferation of Pca cell lines. LNCaP, DU145, PC3 and 22Rv1 cells were treated with 5 nM siControl or siRFC2 (#1, #2, and #3). MTS assay was carried out at the indicated time points (N = 6). The absorbance quantified in the plate reader is shown. The results are presented as mean and SD. One-way ANOVA and Dunnet’s post-doc tests were performed to obtain P-values. * P < 0.05, ** P < 0.01, *** P < 0.001, compared with siControl. ( D ) Knockdown of RFC2 increases DNA damage and apoptosis in Pca cell lines. PC3, DU145 and 22Rv1 cells were transfected with 5 nM siControl or siRFC2 (#1, #2, and #3) for 48 h. Western blot analysis for RFC2, cleaved PARP and γH2AX was carried out and β-actin was used as a loading control.

Article Snippet: The following commercially available antibody was used: rabbit anti-RFC2 (ab251796, Abcam).

Techniques: Inhibition, Expressing, Western Blot, MTS Assay, Transfection

Journal: Scientific Reports

Article Title: Identification of DNA damage response-related genes as biomarkers for castration-resistant prostate cancer

doi: 10.1038/s41598-023-46651-6

Figure Lengend Snippet: Immunohistochemical (IHC) analysis of RFC2 protein expression in Pca tissues. ( A – D ) Representative IHC images of RFC2 in Pca tissues. ( A ) Positive (anti-RFC2 antibody) and negative control (non-specific rabbit IgG antibody) using bladder cancer specimens. ( B ) Anti-RFC2 in benign prostate, intensity score 0. ( C ) Anti-RFC2 in Pca, intensity score 1 and 2. ( D ) Anti-RFC2 in CRPC, intensity score 3. Scale bar = 50 μm. ( E ) Rate of cases in which positive IR was detected by RFC2 IHC in benign (N = 12), Pca (N = 103), and CRPC (N = 15) tissues. Chi-squared test was done to calculate P -value. IR scores of 0‐4 and 5–8 were defined as low and high IR, respectively. ( F , G ) High immunoreactivity (IR) score of RFC2 is associated with poor prognosis of prostate cancer patients. Progression-free survival ( F ) and cancer-specific survival ( G ) of prostate cancer patients are shown (N = 103). Survival curve was obtained by Kaplan–Meier method and P -value was determined by log-rank (Mantel–Cox) test. ( H ) Evaluation of the time to CRPC diagnosis in CRPC cases (N = 15). The median follow-up time from radical prostatectomy to CRPC diagnosis was summarized in CRPC cases with low and high RFC2 IR. Mann–Whitney test was done to calculate P -value.

Article Snippet: The following commercially available antibody was used: rabbit anti-RFC2 (ab251796, Abcam).

Techniques: Immunohistochemistry, Expressing, Negative Control, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Identification of DNA damage response-related genes as biomarkers for castration-resistant prostate cancer

doi: 10.1038/s41598-023-46651-6

Figure Lengend Snippet: Relationship between replication factor C subunit 2 (RFC2) immunoreactivity and clinicopathological findings in human prostate cancer patients (n = 103).

Article Snippet: The following commercially available antibody was used: rabbit anti-RFC2 (ab251796, Abcam).

Techniques:

Journal: Scientific Reports

Article Title: Identification of DNA damage response-related genes as biomarkers for castration-resistant prostate cancer

doi: 10.1038/s41598-023-46651-6

Figure Lengend Snippet: Univariate and multivariate analyses for cancer-specific survival in prostate cancer patients.

Article Snippet: The following commercially available antibody was used: rabbit anti-RFC2 (ab251796, Abcam).

Techniques:

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 1. RFC2 expression levels in pan-cancer. (A) RFC2 expression in cancer tissues compared to normal tissues in Oncomine database. For each pair, red indicated higher expression and blue indicated lower expression. (B) The expression levels of RFC2 in different cancer types from TCGA database were analyzed by TIMER database. (C) The differential expression of RFC2 in tumor tissues from TCGA database was compared with normal tissues from GTEx database. (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing, Quantitative Proteomics

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 2. Comparison of RFC2 expression between tumor and normal tissues (left, red represented tumor tissues while blue represented normal tissues) and corresponding immunohistochemistry images in normal tissues (middle) and tumor tissues (right). (A–F) The differential expression levels of RFC2 in glioblastoma (GBM) tissues (A), diffuse lower-grade gliomas (LGG) tissues (B), cervical cancer (CESC) tissues (C), colon adenocarcinoma (COAD) tissues (D), liver hepatocellular carcinoma (LIHC) tissues (E), lung squamous cell carcinoma (LUSC) tissues (F), and their corresponding normal tissues were evaluated based on GEPIA and HPA databases. (*P < 0.05).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Comparison, Expressing, Immunohistochemistry, Quantitative Proteomics

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 3. The correlation between RFC2 expression and clinicopathological characteristics in primary LGG patients (TCGA cohort, n = 537). (A–G) The correlations of RFC2 expression with LGG clinicopathological features, including age (A), gender (B), WHO grade (C), pathological classification (D), IDH1 mutation status (E), radiation therapy history (F), and seizure history (G). (H) Heatmap showing the association between RFC2 expression and clinicopathological characteristics in LGG patients. (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing, Mutagenesis

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 4. Association between RFC2 expression and overall survival (OS) across cancers in TCGA cohort. (A) Forest plot showing the correlation of RFC2 expression with OS in 33 types of cancer. (B–I) Kaplan–Meier survival analyses displaying the relationships of RFC2 expression with OS in different cancer types, including glioblastoma (GBM) (B), diffuse lower-grade gliomas (LGG) (C), adrenocortical carcinoma (ACC) (D), cervical cancer (CESC) (E), kidney chromophobe (KICH) (F), kidney renal clear cell carcinoma (KIRC) (G), lung adenocarcinoma (LUAD) (H), and ocular melanomas (UVM) (I).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 5. Association between RFC2 expression and disease-specific survival (DSS) across cancers in TCGA cohort. (A) Forest plot showing the correlation of RFC2 expression with DSS in 33 types of cancer. (B–I) Kaplan–Meier survival analyses displaying the relationships of RFC2 expression with DSS in different cancer types, including glioblastoma (GBM) (B), diffuse lower-grade gliomas (LGG) (C), adrenocortical carcinoma (ACC) (D), kidney chromophobe (KICH) (E), kidney renal clear cell carcinoma (KIRC) (F), liver hepatocellular carcinoma (LIHC) (G), lung adenocarcinoma (LUAD) (H), and ocular melanomas (UVM) (I).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 6. Associations of RFC2 expression with disease-free interval (DFI) and progression-free interval (PFI) across cancers in TCGA cohort. (A) Forest plot showing the correlation of RFC2 expression with DFI in 33 types of cancer. (B–D) Kaplan–Meier survival analyses displaying the relationships of RFC2 expression with DFI in different cancer types, including diffuse lower-grade gliomas (LGG) (B), liver hepatocellular carcinoma (LIHC) (C), and prostate Cancer (PRAD) (D). (E) Forest plot showing the correlation of RFC2 expression with PFI in 33 types of cancer. (F–O) Kaplan–Meier survival analyses displaying the relationships of RFC2 expression with PFI in different cancer types, including LGG (F), adrenocortical carcinoma (ACC) (G), kidney chromophobe (KICH) (H), kidney renal clear cell carcinoma (KIRC) (I), LIHC (J), lung adenocarcinoma (LUAD) (K), mesothelioma (MESO) (L), pancreatic cancer (PAAD) (M), prostate adenocarcinoma (PRAD) (N), and ocular melanomas (UVM) (O).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 7. Associations between RFC2 expression and tumor mutation burden (TMB), tumor microsatellite instability (MSI), and mismatch repair (MMR) genes in pan-cancer. (A) Radar chart displaying the association between RFC2 expression and TMB. Numbers in blue text represent the correlation coefficient. (B) Radar chart displaying the association between RFC2 expression and MSI. Numbers in black text represent the correlation coefficient. (C) Heatmap illustrating the relationship between RFC2 expression and MMR genes. For each pair, the top left triangle represents the P-value, and the bottom right triangle represents the correlation coefficient. (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing, Mutagenesis

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 8. The correlations of RFC2 levels with immune cell infiltration and immune checkpoints. (A) The proportions of 22 immune cell types in each sample of LGG from TCGA cohort were estimated by conducting CIBERSORT algorithm. (B) Correlations of RFC2 expression with differential immune cell types expression in LGG. (C,D) Associations of RFC2 expression with immune score (C) and stromal score (D) were examined by performing ESTIMATE algorithm. (E) Heatmap illustrating the relationship between RFC2 expression and immune checkpoint genes expression across 33 types of cancer. For each pair, the bottom left triangle represents the correlation coefficient, and the top right triangle represents the P-value. (F) Association between RFC2 and immune checkpoints genes in LGG. (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 9. The functional enrichment analyses of RFC2 in LGG. (A) Heatmap showing the top 50 genes most positively related to RFC2. (B–D) The top 20 Gene Ontology (GO) terms related to RFC2 in biological processes (BP) categories (B), molecular function (MF) categories (C), and cell component (CC) categories (D) respectively. (E) The top 20 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways correlated with RFC2. (F) The top 20 Reactome pathways correlated with RFC2. (G) GSEA results showing the top 5 KEGG pathways associated with RFC2. (H) GSEA results showing the top 5 HALLMARK pathways associated with RFC2.

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Functional Assay

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 10. RFC2 knockdown inhibited the progression of LGG cells. (A,B) The relative expression levels of RFC2 in LGG cells (HS683 and SW1783) and microglia (HMC3) were estimated by Western blot analysis (A) and qRT-PCR analysis (B). (C) The relative expression levels of RFC2 in LGG cells transfected with sh1-RFC2, sh2-RFC2, and sh-NC were evaluated by Western blot analysis. (D) CCK-8 assay was conducted to assess the proliferation of LGG cells transfected with sh1-RFC2, sh2-RFC2, and sh-NC. (E) Colony formation assay was performed to evaluate the colony formation capability of LGG cells transfected with sh1-RFC2, sh2-RFC2, and sh-NC. (F) Apoptotic rate of LGG cells transfected with sh1-RFC2, sh2-RFC2, and sh-NC was detected by using flow cytometry. (G) Cell cycle distribution of LGG cells transfected with sh1-RFC2, sh2-RFC2, and sh-NC was examined by using flow cytometry. (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Knockdown, Expressing, Western Blot, Quantitative RT-PCR, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry

Journal: Scientific reports

Article Title: RFC2: a prognosis biomarker correlated with the immune signature in diffuse lower-grade gliomas.

doi: 10.1038/s41598-022-06197-5

Figure Lengend Snippet: Figure 11. RFC2 expression correlated with clinicopathological features in our LGG samples. (A) The immunohistochemistry images showing the relative expression of RFC2 in LGG tissues and benign brain tumor tissues (Original magnification, × 400). (B) RFC2 expression levels in LGG tissues (n = 69) and benign brain tumor tissues (n = 10) were evaluated by immunohistochemistry analysis. (C–J) The associations of RFC2 expression with clinicopathological factors, including age (C), gender (D), WHO grade (E), pathological classification (F), IDH1 mutation status (G), radiation therapy history (H), chemotherapy history (I), and seizure history (J). (K) Heatmap displaying the correlation between RFC2 expression and clinicopathological features in our LGG patients. (L,M) Kaplan–Meier survival analyses showing the relationships of RFC2 expression with overall survival (OS) (L) and disease-free survival (DFS) (M) in LGG patients. (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: Proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), which were then incubated overnight at 4 °C with primary antibodies, including RFC2 (1:1000, 10410-1-AP, Proteintech, USA) and β-Tubulin (1:1000, 10094-1-AP, Proteintech, USA).

Techniques: Expressing, Immunohistochemistry, Mutagenesis