Journal: Iranian Journal of Pathology

Article Title: Significance of RCC2, Rac1 and p53 Expression in Breast Infiltrating Ductal Carcinoma; An Immunohistochemical Study

doi: 10.30699/IJP.2024.2014367.3198

Figure Lengend Snippet: Expression of RCC2, Rac1 and P53 in the studied IDC cases

Article Snippet: The primary antibodies included ready-to-use IgG Rabbit monoclonal anti-RCC2 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA1680), anti-Rac1 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA2224), and anti-p53 antibody (Biocare Medical, USA, Catalog Number: IP 298 G10).

Techniques: Expressing

Journal: Iranian Journal of Pathology

Article Title: Significance of RCC2, Rac1 and p53 Expression in Breast Infiltrating Ductal Carcinoma; An Immunohistochemical Study

doi: 10.30699/IJP.2024.2014367.3198

Figure Lengend Snippet: Infiltrating ductal carcinoma showing (A) strong nuclear expression of RCC2 (IHC x200), (B) moderate nuclear expression of RCC2 (IHC x200), (C) mild nuclear expression of RCC2 (IHC x200), (D) negative expression of RCC2 (IHC x200).

Article Snippet: The primary antibodies included ready-to-use IgG Rabbit monoclonal anti-RCC2 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA1680), anti-Rac1 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA2224), and anti-p53 antibody (Biocare Medical, USA, Catalog Number: IP 298 G10).

Techniques: Expressing

Journal: Iranian Journal of Pathology

Article Title: Significance of RCC2, Rac1 and p53 Expression in Breast Infiltrating Ductal Carcinoma; An Immunohistochemical Study

doi: 10.30699/IJP.2024.2014367.3198

Figure Lengend Snippet: Relation between RCC2 low/high expression and the clinicopathological parameters in all the studied breast IDC cases (n= 120)

Article Snippet: The primary antibodies included ready-to-use IgG Rabbit monoclonal anti-RCC2 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA1680), anti-Rac1 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA2224), and anti-p53 antibody (Biocare Medical, USA, Catalog Number: IP 298 G10).

Techniques: Expressing

Journal: Iranian Journal of Pathology

Article Title: Significance of RCC2, Rac1 and p53 Expression in Breast Infiltrating Ductal Carcinoma; An Immunohistochemical Study

doi: 10.30699/IJP.2024.2014367.3198

Figure Lengend Snippet: Infiltrating ductal carcinoma with Her2neu positive status (A) and luminal B subtype (B) showing a significantly higher RCC2 expression. Large tumor size (C) shows a significant association with strong RCC2. Positive correlation between p53 percent and apoptosis (D) Significant relation between positive PR and low p53 expression (E) Strong p53 intensity is associated with N0 and N1 nodal stage (F)

Article Snippet: The primary antibodies included ready-to-use IgG Rabbit monoclonal anti-RCC2 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA1680), anti-Rac1 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA2224), and anti-p53 antibody (Biocare Medical, USA, Catalog Number: IP 298 G10).

Techniques: Expressing

Journal: Iranian Journal of Pathology

Article Title: Significance of RCC2, Rac1 and p53 Expression in Breast Infiltrating Ductal Carcinoma; An Immunohistochemical Study

doi: 10.30699/IJP.2024.2014367.3198

Figure Lengend Snippet: A highly significant direct linear correlation between expression of (A) RCC2 and Rac1 ( P <0.001, r=0.424), (B) RCC2 and p53 ( P =<0.001, r=0.315), and (C) Rac1 and p53 ( P =0.006, r=0.332).

Article Snippet: The primary antibodies included ready-to-use IgG Rabbit monoclonal anti-RCC2 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA1680), anti-Rac1 antibody (Chongqing Biospes Co., LTD, China, Catalog Number: YPA2224), and anti-p53 antibody (Biocare Medical, USA, Catalog Number: IP 298 G10).

Techniques: Expressing

Journal: Nature Communications

Article Title: TD-60 links RalA GTPase function to the CPC in mitosis

doi: 10.1038/ncomms8678

Figure Lengend Snippet: ( a ) Immunoblots of whole HeLa cell extract (WCE) transfected with siRNAs targeting two different regions of human TD-60. Immunoblot for RCC2 (TD-60) and RalA, with β-actin as a loading control. ( b ) Mitotic index of TD-60-depleted cells 48 h after transfection with siRNA is increased from prophase to metaphase. Graphs report the mean±s.e.m. ( n =4). ( c ) Control and TD-60-depleted cells fixed with 4% PFA were stained with DAPI (blue), α-tubulin (red) and aurora B (green). Scale bar, 10 μm. ( d ) The proportion of prometaphase cells with abnormal spindle shapes (in which the poles were partly separated but nascent spindles were confined within the regions containing chromosomes) in TD-60-depleted and control cultures was determined by a blinded observer ( n =4). In this and all subsequent figures, graphs report the mean±s.e.m., with statistical significance assessed using the Mann–Whitney U -test (* P <0.05). ( e ) Both prometaphase and metaphase cells show an increased level of centromeric Aurora B after 48 h TD-60 depletion. TD-60-depleted cells were fixed with 4% PFA and stained with DAPI (blue), ACA (red) and Aurora B (green). Scale bar, 10 μm. ( f ) Quantification of Aurora B intensity from the experiment illustrated in e using ImageJ and normalized to the ACA control ( n =15, ** P <0.005 by Mann–Whitney U -test). More information regarding quantification can be found in . DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: Immunoblotting analysis was performed using the following primary antibodies: anti-RCC2 rabbit polyclonal antibody (Cell Signalling 1:100–1:200), anti-RalA mouse monoclonal antibody (BD Bioscience 1:100), anti-Ran mouse monoclonal antibody (BD Bioscience 1:1,000), anti-Tubulin B512 mouse monoclonal antibody (Sigma 1:10,000), anti-Beta Actin mouse monoclonal antibody (Sigma 1:1,000) and anti-GFP rabbit polyclonal (Life Technology, 1 μg ml −1 ).

Techniques: Western Blot, Transfection, Control, Staining, MANN-WHITNEY

Journal: BMC Cancer

Article Title: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

doi: 10.1186/s12885-017-3908-y

Figure Lengend Snippet: Tumor cells expressing RCC2-YFP are resistant to apoptosis. a Tumor cell lines HeLa, MDA-MB-231 and CRL5800 were transfected with a RCC2-YFP expression plasmid or a control YFP plasmid for 24 h, 48 h and 72 h. Western blot analysis with a RCC2-specific antibody showed both RCC2-YFP and endogenous RCC2 in these cells. b HeLa cells were cultured in 96 well plates for 1–3 days and counted by trypan blue stain. Cells expressing RCC2-YFP had increased cell proliferation compared to control YFP HeLa cells or parental cells (mean ± S.D. of four replicates, P < 0.05 at day 3). c HeLa cells were cultured in soft agar plates for 10 days. RCC2-YFP #1 and #2 were cells from two independent transfections. Bigger cell colonies were seen in the RCC2-YFP-expressing cells as compared to the YFP-expressing cells. d & e HeLa cells were transiently transfected with YFP or RCC2-YFP for 24 h, fixed and counterstained with DAPI. Apoptotic cells were scored by nucleus pyknosis and fragmentation (arrows). RCC2-YFP expression virtually eliminated spontaneous apoptosis in both serum-free and 10% FBS culture conditions (mean ± S.D. of four replicates, *P < 0.01) ( e ). f MDA-MB-231 and CRL5800 cells were transiently transfected with YFP or RCC2-YFP, treated with 10 μM STS for 48 h and apoptotic cells scored by DAPI stain. RCC2-YFP expression partially blocked the STS-induced apoptosis (mean ± S.D. of four replicates, *P < 0.01)

Article Snippet: The anti-RCC2 antibody (D14F3, cell signaling) was validated by IHC of cultured HeLa cells, which showed a nuclear localization in interphase cells and spindle midzone/midbody localization in mitosis as expected (Fig. ).

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Western Blot, Cell Culture, Staining

Journal: BMC Cancer

Article Title: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

doi: 10.1186/s12885-017-3908-y

Figure Lengend Snippet: RCC2 down regulation in tumor cells led to increased sensitivity to STS treatment. a & b Tumor cell lines HeLa, MDA-MB-231 and CRL5800 were transfected with 3 RCC2-specific siRNA oligos (siRNA-RCC2–1, siRNA-RCC2–2, siRNA-RCC2–3) or a control siRNA (siNC) for 24 h, 48 h, and 72 h. RCC2 protein ( a ) and mRNA ( b ) were evaluated by Western blotting and quantitative real-time RT-PCR respectively. All 3 RCC2-specific siRNAs were effective in reducing RCC2 expression (mean ± S.D. of four replicates, *P < 0.01). c Tumor cells transfected with siRNA-RCC2–1, siRNA-RCC2–2, or siNC for 24 h were treated with STS at increasing concentrations (0.05, 0.1, 0.2 and 0.4 μM for HeLa; 5, 8, 10, 20 μM for MDA-MB-231; 5, 10, 30 and 50 μM for CRL5800). Surviving cells were estimated by a CellTiter-Glo® Luminescent Cell Viability Assay. Luminescence values represent the mean ± S.D. of six wells, *P < 0.05)

Article Snippet: The anti-RCC2 antibody (D14F3, cell signaling) was validated by IHC of cultured HeLa cells, which showed a nuclear localization in interphase cells and spindle midzone/midbody localization in mitosis as expected (Fig. ).

Techniques: Transfection, Control, Western Blot, Quantitative RT-PCR, Expressing, Cell Viability Assay

Journal: BMC Cancer

Article Title: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

doi: 10.1186/s12885-017-3908-y

Figure Lengend Snippet: RCC2-YFP interrupts apoptosis via blocking Rac1 activation. a Protein lysate from HeLa cells expressing YFP or RCC2-YFP were immunoprecipitated with anti-GFP antibody and Western blotted with anti-GFP, anti-Rac1, anti-cdc42, or anti-RhoA antibody. Rac1 was co-precipitated with RCC2-YFP. b HeLa cells expressing YFP or RCC2-YFP were serum-starve1d overnight followed by serum-stimulation for 5 min. GTP-bound activated Rac1 was pulled down by PAK-PBD beads and Western blotted with anti-Rac1 antibody. RCC2-YFP expression blocked both endogenous and serum-induced Rac1 activation. c HeLa, MDA-MA-231 and CRL5800 cells expressing YFP or RCC2-YFP were treated with STS (0.5 μM for HeLa; 10 μM for MDA and CRL5800) for 30 min and Western blotted with antibodies to phospho-JNK and total JNK. RCC2-YFP expression blocked the STS-induced JNK phosphorylation in all three cell lines. d Tumor cells transfected with Rac1-Q61L, RCC2-YFP or both for 48 h were treated with STS (0.5 μM for HeLa; 10 μM for MDA and CRL5800) and apoptotic cells were scored by DAPI stain after 24 h treatment. Rac1-Q61L expression in tumor cells revoked the protection of RCC2-YFP towards STS-induced apoptosis (mean ± S.D. of four replicates, *P < 0.05). e HeLa cells transfected with Rac1-Q61L, RCC2-YFP or both for 48 h in the presence of serum. Spontaneous apoptotic cells were scored by DAPI stain. Rac1-Q61L expression in HeLa cells revoked the protection of RCC2-YFP toward spontaneous apoptosis (mean ± S.D. of four replicates, *P < 0.01). f HeLa cells were transfected with Rac1-Q61L, RCC2-YFP or both in the presence of serum, and Caspase-Glo® 3/7 assay was performed 48 h after transfection. Rac1-Q61L and RCC2-YFP expression in HeLa cells affected Caspase 3/7 activity (mean ± S.D. of six wells, *P < 0.05)

Article Snippet: The anti-RCC2 antibody (D14F3, cell signaling) was validated by IHC of cultured HeLa cells, which showed a nuclear localization in interphase cells and spindle midzone/midbody localization in mitosis as expected (Fig. ).

Techniques: Blocking Assay, Activation Assay, Expressing, Immunoprecipitation, Western Blot, Phospho-proteomics, Transfection, Staining, Caspase-Glo Assay, Activity Assay

Journal: BMC Cancer

Article Title: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

doi: 10.1186/s12885-017-3908-y

Figure Lengend Snippet: RCC2-YFP expression in tumor cells affects drug sensitivity. a HeLa, CRL5800 and MDA-MA-231 cells expressing YFP or RCC2-YFP were treated with vehicles or increasing doses of Taxol, Nocodazole (Noco), Hydroxyurea (Hydrea), Daunorubicin (Dauno), Cisplatin, Camptothecin (CPT), Staurosporine (STS), 5-Fluorouracil (5-FU) or Irinotecan for 48 h. b, c, d and e Cells were quantitated by a CellTiter-Glo® Luminescent Cell Viability Assay. No significant difference was observed between untreated cells and vehicle-treated cells (0.1% DMSO or 0.1% saline) ( b ). Surviving cells were calculated as the fraction of vehicle controls ( c, d and e ). Results were averaged between six wells per dose in two experiments (mean ± S.D.)

Article Snippet: The anti-RCC2 antibody (D14F3, cell signaling) was validated by IHC of cultured HeLa cells, which showed a nuclear localization in interphase cells and spindle midzone/midbody localization in mitosis as expected (Fig. ).

Techniques: Expressing, Cell Viability Assay, Saline

Journal: BMC Cancer

Article Title: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

doi: 10.1186/s12885-017-3908-y

Figure Lengend Snippet: RCC2 was over-expressed in lung cancers and ovarian cancers. a IHC of HeLa cells was performed with anti-RCC2 antibody (D14F3, cell signaling). RCC2 signals were seen in nucleus and midbody of a telophase cell as expected. Cytoplasmic RCC2 was also observed. b RCC2 expression in a lung cancer tissue microarray was evaluated by IHC. RCC2 were not seen in normal lung (left) but highly expressed in lung cancer (10×, scale bar: 100 μm). In addition to nuclei, RCC2 signals were also seen in cytoplasm in some lung cancers ( c ). d RCC2 expression in an ovarian cancer tissue microarray was evaluated by IHC. Normal ovaries expressed none or weak RCC2 (−~+) (left). Increased RCC2 expression was seen in majority of ovarian cancers

Article Snippet: The anti-RCC2 antibody (D14F3, cell signaling) was validated by IHC of cultured HeLa cells, which showed a nuclear localization in interphase cells and spindle midzone/midbody localization in mitosis as expected (Fig. ).

Techniques: Expressing, Microarray

Journal: BMC Cancer

Article Title: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

doi: 10.1186/s12885-017-3908-y

Figure Lengend Snippet: Correlation between RCC2 expression and clinicopathological features in lung cancer

Article Snippet: The anti-RCC2 antibody (D14F3, cell signaling) was validated by IHC of cultured HeLa cells, which showed a nuclear localization in interphase cells and spindle midzone/midbody localization in mitosis as expected (Fig. ).

Techniques: Expressing

Journal: BMC Cancer

Article Title: RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation

doi: 10.1186/s12885-017-3908-y

Figure Lengend Snippet: Correlation between RCC2 expression and clinicopathological features in ovarian cancer

Article Snippet: The anti-RCC2 antibody (D14F3, cell signaling) was validated by IHC of cultured HeLa cells, which showed a nuclear localization in interphase cells and spindle midzone/midbody localization in mitosis as expected (Fig. ).

Techniques: Expressing