Journal: Journal of Nanobiotechnology

Article Title: Zinc oxide nanosphere for hydrogen sulfide scavenging and ferroptosis of colorectal cancer

doi: 10.1186/s12951-021-01069-y

Figure Lengend Snippet: Transcriptome analysis in VZnO-treated HCT116. A KEGG pathway analysis based on the RNA-seq results in HCT116. B Representative heatmap of gene expression levels. C Representative scatter plot of 140 significant genes (33 unregulated genes marked in red and 107 downregulated genes marked in green) for Treat vs. Con. D mRNA levels of GGCT, RRM1 and RRM2 by RNA-seq. E Representative Western Blot result of GGCT, RRM1 and RRM2. Membranes were re-probed for GAPDH expression to show that similar amounts of protein were loaded in each lane

Article Snippet: Primary antibodies were used at the following dilutions: rabbit-anti-GPX4 (Sigma-SAB4300725) 1:2000; rabbit-anti-CBS (Sigma-AV45746) 1:4000; rabbit-anti-COX2 (Sigma-SAB4200576) 1:5000; goat-anti-NOX1 (Sigma-SAB2501686) 1:5000; rabbit-anti-TRF1 (Sigma-SAB4502943) 1:500; rabbit-anti-GAPDH (Invitrogen-PA1-987) 1:3000; rabbit-anti-GGCT (Invitrogen-PA5-80,658) 1:1000; rabbit-anti-RRM1 (Invitrogen-PA5-75,989) 1:1000; rabbit-anti-RRM2 (Invitrogen-PA5-27,856) 1:1000.

Techniques: RNA Sequencing Assay, Expressing, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: Age-associated changes in microglia and astrocytes ameliorate blood-brain barrier dysfunction

doi: 10.1016/j.omtn.2021.08.030

Figure Lengend Snippet: Primers used in this study

Article Snippet: The following antibodies were used for western blot: rabbit anti-DNAJA1 (Proteintech, 11713-1-AP); rabbit anti-DNAJB1 (Proteintech, 13174-1-AP); rabbit anti-HSPH1 (Proteintech, 13383-1-AP); rabbit anti-RRM2 (Invitrogen, PA5-79937); rabbit anti-HSPA1A (Invitrogen, PA5-34772); rabbit anti-HSPA1B (Invitrogen, PA5-97846); mouse anti-dCK (Santa Cruz Biotechnology, sc-393099); mouse anti-Nr4a1 (Santa Cruz Biotechnology, sc-365113); and rabbit anti-GAPDH (Cell Signaling Technology, 2118S).

Techniques:

Journal: BMC Cancer

Article Title: RRM2 and CDC6 are novel effectors of XBP1-mediated endocrine resistance and predictive markers of tamoxifen sensitivity

doi: 10.1186/s12885-023-10745-1

Figure Lengend Snippet: XBP1 is required for expression of RRM2, CDC6 and TOP2A. A Total RNA and whole cell lysate from control and XBP1 KO MCF7 cells was used to determine the expression of indicated genes. Expression level of indicated genes was evaluated by qRT-PCR and normalised against RPLP0. Data presented is mean ± S.D ( n = 3). * p < 0.05, ** p < 0.01, two-tailed unpaired t-test compared with control cells. Right panel, a representative immunoblot for RRM2, CDC6, TOP2 and β-Actin is shown ( n = 3). B-D MCF7, T47D and BT474 cells were treated STF083010 (50 µM) for 96 h. Total RNA and whole cell lysate was used to determine the expression of indicated genes. Expression level of indicated genes was determined by real time RT-PCR and normalised against RPLP0. Data presented as mean ± S.D ( n = 3). A representative immunoblot and quantification of RRM2, CDC6, TOP2A expression normalised to β-Actin is shown ( n = 3)

Article Snippet: Mouse ER-α Ab (Cat# sc-8002), Mouse RRM2 Ab (sc-376973), mouse CDC6 Ab (Cat# sc-9964) were purchased from Santa Cruz Biotechnology, Inc. (Bergheimer, Heidelberg, Germany).

Techniques: Expressing, Control, Quantitative RT-PCR, Two Tailed Test, Western Blot

Journal: BMC Cancer

Article Title: RRM2 and CDC6 are novel effectors of XBP1-mediated endocrine resistance and predictive markers of tamoxifen sensitivity

doi: 10.1186/s12885-023-10745-1

Figure Lengend Snippet: Ligand independent induction of RRM2, CDC6 and TOP2A in ESR1 mutant MCF7 cells. MCF7 WT, MCF7 Y537S, MCF7 D538G cells were synchronized for 3 days in phenol red free DMEM containing 5% CSS. MCF7 WT cells were either treated with (Veh) or estrogen (E2) for 24 h. Expression level of RRM2, CDC6, and TOP2A was evaluated by qRT-PCR and normalised against RPLP0. Data presented as mean ± S.D of three independent experiments. * p < 0.05, two-tailed unpaired t-test compared with vehicle treated MCF7 WT cells

Article Snippet: Mouse ER-α Ab (Cat# sc-8002), Mouse RRM2 Ab (sc-376973), mouse CDC6 Ab (Cat# sc-9964) were purchased from Santa Cruz Biotechnology, Inc. (Bergheimer, Heidelberg, Germany).

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

Journal: BMC Cancer

Article Title: RRM2 and CDC6 are novel effectors of XBP1-mediated endocrine resistance and predictive markers of tamoxifen sensitivity

doi: 10.1186/s12885-023-10745-1

Figure Lengend Snippet: Expression of RRM2 and CDC6 rescues tamoxifen hypersensitivity in XBP1 knockout MCF7 cells. A Whole cell lysates from indicated RRM2 expressing cells were analysed by western blotting using antibodies against RRM2 and β-Actin. B Indicated RRM2 expressing cells were treated with tamoxifen (10 µM) for required time points. Line graphs show change in O.D at 490 nm. Data presented is mean ± S.D ( n = 4). * p < 0.05, two-tailed unpaired t-test comparing respective time points. C Whole cell lysates from indicated (wild type and mutant) CDC6 expressing cells were subjected to western blotting using antibodies against CDC6 and β-Actin. D Indicated mutant CDC6 expressing cells were treated with tamoxifen (10 µM) for required time points. Line graphs show the change in O.D at 490 nm. Data presented as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01 from two-tailed unpaired t-test comparing respective time points

Article Snippet: Mouse ER-α Ab (Cat# sc-8002), Mouse RRM2 Ab (sc-376973), mouse CDC6 Ab (Cat# sc-9964) were purchased from Santa Cruz Biotechnology, Inc. (Bergheimer, Heidelberg, Germany).

Techniques: Expressing, Knock-Out, Western Blot, Two Tailed Test, Mutagenesis

Journal: BMC Cancer

Article Title: RRM2 and CDC6 are novel effectors of XBP1-mediated endocrine resistance and predictive markers of tamoxifen sensitivity

doi: 10.1186/s12885-023-10745-1

Figure Lengend Snippet: Association of XBP1-gene signature with outcome in ER-positive breast cancer. A Box plot for expression of RRM2, CDC6 and TOP2A in tumour and normal tissues in human breast cancers is shown. Median is shown by horizontal black line, the box is the upper and lower quartiles and the two lines outside the box show the highest and lowest values. B KM Plotter ( https://kmplot.com/analysis/ ) was used to determine the association of XBP1-gene signature (CDC6, RRM2 and TOP2A) with overall survival (OS) and recurrence free survival (RFS) in ER-positive breast cancer. C Web-based algorithm PROGgeneV 2 ( http://www.progtools.net/gene/index.php ) was used to test the association between XBP1-gene signature and RFS in indicated datasets of breast cancer. D Web-based algorithm ROC plot ( http://www.rocplot.org/ ) was used to evaluate the association between XBP1-gene signature and response to tamoxifen treatment in ER-positive breast cancer

Article Snippet: Mouse ER-α Ab (Cat# sc-8002), Mouse RRM2 Ab (sc-376973), mouse CDC6 Ab (Cat# sc-9964) were purchased from Santa Cruz Biotechnology, Inc. (Bergheimer, Heidelberg, Germany).

Techniques: Expressing

Journal: BMC Cancer

Article Title: RRM2 and CDC6 are novel effectors of XBP1-mediated endocrine resistance and predictive markers of tamoxifen sensitivity

doi: 10.1186/s12885-023-10745-1

Figure Lengend Snippet: Graphical summary. Hypoxia and glucose deprivation are physiologically important inducers of unfolded protein response in tumour microenvironment. In estrogen receptor (ER) positive breast cancer cells XBP1s is induced in response to E2-stimulation and conditions of UPR. Tumour cells survive stressful conditions of microenvironment by an adaptive mechanism called the unfolded protein response (UPR). XBP1s is a transcriptional activator that regulates expression of genes involved in protein homeostasis and promote cell survival during conditions of UPR. XBP1s is required for optimal induction of E2-responsive genes and RRM2, CDC6 and TOP2A. RRM2 and CDC6 mediate endocrine resistance downstream of XBP1s in ER-positive breast cancer. ER, Estrogen receptor alpha; XBP1, X-box binding protein 1

Article Snippet: Mouse ER-α Ab (Cat# sc-8002), Mouse RRM2 Ab (sc-376973), mouse CDC6 Ab (Cat# sc-9964) were purchased from Santa Cruz Biotechnology, Inc. (Bergheimer, Heidelberg, Germany).

Techniques: Expressing, Binding Assay