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stage iv metastatic biopsies  (OriGene)


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    OriGene stage iv metastatic biopsies
    ( A to C ) Lung colonization assay for control and either RBMS3 knockdown or RBMS3 overexpression. Luciferase-labeled cells were injected via tail vein, and their <t>metastatic</t> growth in the lungs was measured over time. The magnitude of the signal, shown here as a heatmap for a representative mouse from each cohort, reflects the metastatic burden. Bars on time-course plots show means ± SEM, and ANOVA was performed (left). Also included are area under the curve (AUC) of log-normalized signal in the lungs of mice over 25 days. Summary overlay shows means ± SEM and Mann-Whitney U test used to determine statistical significance (middle). Also, hematoxylin and eosin–stained lung sections for exemplary mice (right). (A) Lung colonization assays of control ( N = 4 biological replicates) and RBMS3 knockdown MDA-MB-231 cells ( N = 4 biological replicates). (B) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 overexpression MDA-LM2 cells ( N = 5 biological replicates). (C) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 knockdown HCC1806 cells ( N = 5 biological replicates). ( D ) Invasion assays for control and RBMS3 knockdown MDA-MB-231 cells. Images are representative (i.e., median) in each group. In processed images, black is empty space and white is cells. The graph shows fraction that is white (i.e., % area covered), and the Mann-Whitney U test was used for statistical comparison. * P < 0.05; *** P < 0.001.
    Stage Iv Metastatic Biopsies, supplied by OriGene, used in various techniques. Bioz Stars score: 94/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/breast+cancer+cdna+array+ii/TissueScan%2C+Breast+Cancer+cDNA+Array+II/pmc12978252-415-31-35
    Average 94 stars, based on 39 article reviews
    stage iv metastatic biopsies - by Bioz Stars, 2026-10
    94/100 stars

    Images

    1) Product Images from "Integrative analysis of mRNA stability regulation uncovers a metastasis-suppressive program in breast cancer"

    Article Title: Integrative analysis of mRNA stability regulation uncovers a metastasis-suppressive program in breast cancer

    Journal: Science Advances

    doi: 10.1126/sciadv.aea9061

    ( A to C ) Lung colonization assay for control and either RBMS3 knockdown or RBMS3 overexpression. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. The magnitude of the signal, shown here as a heatmap for a representative mouse from each cohort, reflects the metastatic burden. Bars on time-course plots show means ± SEM, and ANOVA was performed (left). Also included are area under the curve (AUC) of log-normalized signal in the lungs of mice over 25 days. Summary overlay shows means ± SEM and Mann-Whitney U test used to determine statistical significance (middle). Also, hematoxylin and eosin–stained lung sections for exemplary mice (right). (A) Lung colonization assays of control ( N = 4 biological replicates) and RBMS3 knockdown MDA-MB-231 cells ( N = 4 biological replicates). (B) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 overexpression MDA-LM2 cells ( N = 5 biological replicates). (C) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 knockdown HCC1806 cells ( N = 5 biological replicates). ( D ) Invasion assays for control and RBMS3 knockdown MDA-MB-231 cells. Images are representative (i.e., median) in each group. In processed images, black is empty space and white is cells. The graph shows fraction that is white (i.e., % area covered), and the Mann-Whitney U test was used for statistical comparison. * P < 0.05; *** P < 0.001.
    Figure Legend Snippet: ( A to C ) Lung colonization assay for control and either RBMS3 knockdown or RBMS3 overexpression. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. The magnitude of the signal, shown here as a heatmap for a representative mouse from each cohort, reflects the metastatic burden. Bars on time-course plots show means ± SEM, and ANOVA was performed (left). Also included are area under the curve (AUC) of log-normalized signal in the lungs of mice over 25 days. Summary overlay shows means ± SEM and Mann-Whitney U test used to determine statistical significance (middle). Also, hematoxylin and eosin–stained lung sections for exemplary mice (right). (A) Lung colonization assays of control ( N = 4 biological replicates) and RBMS3 knockdown MDA-MB-231 cells ( N = 4 biological replicates). (B) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 overexpression MDA-LM2 cells ( N = 5 biological replicates). (C) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 knockdown HCC1806 cells ( N = 5 biological replicates). ( D ) Invasion assays for control and RBMS3 knockdown MDA-MB-231 cells. Images are representative (i.e., median) in each group. In processed images, black is empty space and white is cells. The graph shows fraction that is white (i.e., % area covered), and the Mann-Whitney U test was used for statistical comparison. * P < 0.05; *** P < 0.001.

    Techniques Used: Control, Knockdown, Over Expression, Luciferase, Labeling, Injection, MANN-WHITNEY, Staining, Comparison

    ( A ) Schematic of the dual-guide CRISPR interference (CRISPRi) screen. ( B ) Analysis of CRISPRi screen comparing the abundance of cells expressing each guide between in vivo– and in vitro–grown cells with DESeq2. TXNIP was observed to be present at high levels in vivo and low levels in vitro indicating high metastasis but low proliferation. ( C ) Comparative analysis of TXNIP to RBMS3 expression in the METABRIC cohort . Shown is Pearson correlation with t statistic for nonzero correlation. ( D ) Analysis of disease-free survival in the METABRIC cohort relative to TXNIP expression. ( E ) Meta-analysis of relapse-free survival in smaller published cohorts relative to TXNIP expression. (D and E) The Mantel-Cox test was used to measure significance. Low TXNIP expression is indicative of poor prognosis for disease-free survival in patients with breast cancer. ( F ) The expression levels of TXNIP in 90 tumor samples at different stages of breast cancer was measured by qPCR; bars show means and SEM. ANOVA was performed. ( G ) The expression levels of TXNIP in RBMS3 knockdown MDA-MB-231 cells and control cells was measured by qPCR and compared using Mann-Whitney U test. ( H ) Lung colonization assays of control ( N = 5 mice), TXNIP knockdown ( N = 5 mice), and RBMS3-TXNIP double knockdown ( N = 5 mice) in MDA-MB-231 cells. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. ANOVA was performed; bars indicate means and SEM. ( I ) Molecular mechanism of RBMS3 metastasis suppression through posttranscriptional regulatory action.
    Figure Legend Snippet: ( A ) Schematic of the dual-guide CRISPR interference (CRISPRi) screen. ( B ) Analysis of CRISPRi screen comparing the abundance of cells expressing each guide between in vivo– and in vitro–grown cells with DESeq2. TXNIP was observed to be present at high levels in vivo and low levels in vitro indicating high metastasis but low proliferation. ( C ) Comparative analysis of TXNIP to RBMS3 expression in the METABRIC cohort . Shown is Pearson correlation with t statistic for nonzero correlation. ( D ) Analysis of disease-free survival in the METABRIC cohort relative to TXNIP expression. ( E ) Meta-analysis of relapse-free survival in smaller published cohorts relative to TXNIP expression. (D and E) The Mantel-Cox test was used to measure significance. Low TXNIP expression is indicative of poor prognosis for disease-free survival in patients with breast cancer. ( F ) The expression levels of TXNIP in 90 tumor samples at different stages of breast cancer was measured by qPCR; bars show means and SEM. ANOVA was performed. ( G ) The expression levels of TXNIP in RBMS3 knockdown MDA-MB-231 cells and control cells was measured by qPCR and compared using Mann-Whitney U test. ( H ) Lung colonization assays of control ( N = 5 mice), TXNIP knockdown ( N = 5 mice), and RBMS3-TXNIP double knockdown ( N = 5 mice) in MDA-MB-231 cells. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. ANOVA was performed; bars indicate means and SEM. ( I ) Molecular mechanism of RBMS3 metastasis suppression through posttranscriptional regulatory action.

    Techniques Used: CRISPR, Expressing, In Vivo, In Vitro, Knockdown, Control, MANN-WHITNEY, Luciferase, Labeling, Injection

    Related Articles

    other:

    Article Title: Microfluidic source-sink model reveals effects of biophysically distinct CXCL12 isoforms in breast cancer chemotaxis.
    Article Snippet: We used a cDNA array of normal human breast tissue and breast cancers (Origene, Breast Cancer cDNA Array II).

    Real-time Polymerase Chain Reaction:

    Article Title: The a3 isoform of subunit a of the vacuolar ATPase localizes to the plasma membrane of invasive breast tumor cells and is overexpressed in human breast cancer.
    Article Snippet: Cells that had migrated to the trans-membrane were imaged using a Zeiss Axiovert 10 fluorescence microscope, with an average of 10 images taken per well. .. The number of migrating cells was averaged over three wells per condition with all experiments performed at least three times. qPCR The breast cancer cDNA array II was obtained from Origene. .. The cDNA array contained cDNA from 5 normal breast tissues and 43 breast tumor tissues of varying stage and receptor status. qPCR was performed using primers for beta-actin (supplied by Origene), human subunit a3 (forward: 5′-GACCGACAGGAGGAAAACAAG-3′, reverse: 5′-GT AGGAGGCGGTGTTGGA-3′), or human subunit a4 (forward: 5′- GGAAATCCCAGCTGCAGGC-3′, reverse: 5′- GCAGCCCAGGCAGTACTCG-3′) according to the manufacturer’s instructions using SYBR Green qPCR master mix.

    Software:

    Article Title: Dickkopf-1 is regulated by the mevalonate pathway in breast cancer.
    Article Snippet: .. Assessment of three individual experiments was conducted using the software provided by the manufacturer. cDNA polymerase chain reaction array The breast cancer cDNA Array II was purchased from Origene (Rockville, MD, USA) and was assessed for DKK-1 expression, normalized to GAPDH using the supplier’s protocol. ..

    Polymerase Chain Reaction:

    Article Title: Dickkopf-1 is regulated by the mevalonate pathway in breast cancer.
    Article Snippet: .. Assessment of three individual experiments was conducted using the software provided by the manufacturer. cDNA polymerase chain reaction array The breast cancer cDNA Array II was purchased from Origene (Rockville, MD, USA) and was assessed for DKK-1 expression, normalized to GAPDH using the supplier’s protocol. ..

    Expressing:

    Article Title: Dickkopf-1 is regulated by the mevalonate pathway in breast cancer.
    Article Snippet: .. Assessment of three individual experiments was conducted using the software provided by the manufacturer. cDNA polymerase chain reaction array The breast cancer cDNA Array II was purchased from Origene (Rockville, MD, USA) and was assessed for DKK-1 expression, normalized to GAPDH using the supplier’s protocol. ..

    Article Title: The expression signature of cancer-associated KRAB-ZNF factors identified in TCGA pan-cancer transcriptomic data.
    Article Snippet: .. To test KRAB-ZNF expression in breast cancer, we used commercially available Breast Cancer cDNA Array II purchased from Origene (Rockville, MD, USA) that also provided the clinical and molecular information of patients from whom samples were collected. ..

    Article Title: The expression signature of cancer‐associated KRAB ‐ ZNF factors identified in TCGA pan‐cancer transcriptomic data
    Article Snippet: .. To test KRAB‐ZNF expression in breast cancer, we used commercially available Breast Cancer cDNA Array II purchased from Origene (Rockville, MD, USA) that also provided the clinical and molecular information of patients from whom samples were collected. ..

    Article Title: Dickkopf-1 is regulated by the mevalonate pathway in breast cancer
    Article Snippet: .. The breast cancer cDNA Array II was purchased from Origene (Rockville, MD, USA) and was assessed for DKK-1 expression, normalized to GAPDH using the supplier’s protocol. ..

    Article Title: Estrone-3-Sulfate Stimulates the Proliferation of T47D Breast Cancer Cells Stably Transfected With the Sodium-Dependent Organic Anion Transporter SOAT (SLC10A6)
    Article Snippet: TissueScan TM breast cancer cDNA arrays I-IV (BCRT101-BCRT104) were obtained from OriGene (Rockville, MD, United States). .. In order to analyze SOAT expression in breast cancer, the following TissueScan TM cDNA arrays were commercially obtained from OriGene (Rockville, MD, United States): Breast Cancer cDNA Array I (BCRT101), Breast Cancer cDNA Array II (BCRT102), Breast Cancer cDNA Array III (BCRT103), and Breast Cancer cDNA Array IV (BCRT104). ..



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    ( A to C ) Lung colonization assay for control and either RBMS3 knockdown or RBMS3 overexpression. Luciferase-labeled cells were injected via tail vein, and their <t>metastatic</t> growth in the lungs was measured over time. The magnitude of the signal, shown here as a heatmap for a representative mouse from each cohort, reflects the metastatic burden. Bars on time-course plots show means ± SEM, and ANOVA was performed (left). Also included are area under the curve (AUC) of log-normalized signal in the lungs of mice over 25 days. Summary overlay shows means ± SEM and Mann-Whitney U test used to determine statistical significance (middle). Also, hematoxylin and eosin–stained lung sections for exemplary mice (right). (A) Lung colonization assays of control ( N = 4 biological replicates) and RBMS3 knockdown MDA-MB-231 cells ( N = 4 biological replicates). (B) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 overexpression MDA-LM2 cells ( N = 5 biological replicates). (C) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 knockdown HCC1806 cells ( N = 5 biological replicates). ( D ) Invasion assays for control and RBMS3 knockdown MDA-MB-231 cells. Images are representative (i.e., median) in each group. In processed images, black is empty space and white is cells. The graph shows fraction that is white (i.e., % area covered), and the Mann-Whitney U test was used for statistical comparison. * P < 0.05; *** P < 0.001.
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    Image Search Results


    ( A to C ) Lung colonization assay for control and either RBMS3 knockdown or RBMS3 overexpression. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. The magnitude of the signal, shown here as a heatmap for a representative mouse from each cohort, reflects the metastatic burden. Bars on time-course plots show means ± SEM, and ANOVA was performed (left). Also included are area under the curve (AUC) of log-normalized signal in the lungs of mice over 25 days. Summary overlay shows means ± SEM and Mann-Whitney U test used to determine statistical significance (middle). Also, hematoxylin and eosin–stained lung sections for exemplary mice (right). (A) Lung colonization assays of control ( N = 4 biological replicates) and RBMS3 knockdown MDA-MB-231 cells ( N = 4 biological replicates). (B) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 overexpression MDA-LM2 cells ( N = 5 biological replicates). (C) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 knockdown HCC1806 cells ( N = 5 biological replicates). ( D ) Invasion assays for control and RBMS3 knockdown MDA-MB-231 cells. Images are representative (i.e., median) in each group. In processed images, black is empty space and white is cells. The graph shows fraction that is white (i.e., % area covered), and the Mann-Whitney U test was used for statistical comparison. * P < 0.05; *** P < 0.001.

    Journal: Science Advances

    Article Title: Integrative analysis of mRNA stability regulation uncovers a metastasis-suppressive program in breast cancer

    doi: 10.1126/sciadv.aea9061

    Figure Lengend Snippet: ( A to C ) Lung colonization assay for control and either RBMS3 knockdown or RBMS3 overexpression. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. The magnitude of the signal, shown here as a heatmap for a representative mouse from each cohort, reflects the metastatic burden. Bars on time-course plots show means ± SEM, and ANOVA was performed (left). Also included are area under the curve (AUC) of log-normalized signal in the lungs of mice over 25 days. Summary overlay shows means ± SEM and Mann-Whitney U test used to determine statistical significance (middle). Also, hematoxylin and eosin–stained lung sections for exemplary mice (right). (A) Lung colonization assays of control ( N = 4 biological replicates) and RBMS3 knockdown MDA-MB-231 cells ( N = 4 biological replicates). (B) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 overexpression MDA-LM2 cells ( N = 5 biological replicates). (C) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 knockdown HCC1806 cells ( N = 5 biological replicates). ( D ) Invasion assays for control and RBMS3 knockdown MDA-MB-231 cells. Images are representative (i.e., median) in each group. In processed images, black is empty space and white is cells. The graph shows fraction that is white (i.e., % area covered), and the Mann-Whitney U test was used for statistical comparison. * P < 0.05; *** P < 0.001.

    Article Snippet: We measured TXNIP expression using qPCR in 96 clinical samples across all stages of breast cancer, namely 5 normal epithelial, 23 stage I, 30 stage II, 29 stage III, and 9 stage IV metastatic biopsies (Origene, BCRT102, BCRT103), from which 90 samples yielded sufficient amount of cDNA.

    Techniques: Control, Knockdown, Over Expression, Luciferase, Labeling, Injection, MANN-WHITNEY, Staining, Comparison

    ( A ) Schematic of the dual-guide CRISPR interference (CRISPRi) screen. ( B ) Analysis of CRISPRi screen comparing the abundance of cells expressing each guide between in vivo– and in vitro–grown cells with DESeq2. TXNIP was observed to be present at high levels in vivo and low levels in vitro indicating high metastasis but low proliferation. ( C ) Comparative analysis of TXNIP to RBMS3 expression in the METABRIC cohort . Shown is Pearson correlation with t statistic for nonzero correlation. ( D ) Analysis of disease-free survival in the METABRIC cohort relative to TXNIP expression. ( E ) Meta-analysis of relapse-free survival in smaller published cohorts relative to TXNIP expression. (D and E) The Mantel-Cox test was used to measure significance. Low TXNIP expression is indicative of poor prognosis for disease-free survival in patients with breast cancer. ( F ) The expression levels of TXNIP in 90 tumor samples at different stages of breast cancer was measured by qPCR; bars show means and SEM. ANOVA was performed. ( G ) The expression levels of TXNIP in RBMS3 knockdown MDA-MB-231 cells and control cells was measured by qPCR and compared using Mann-Whitney U test. ( H ) Lung colonization assays of control ( N = 5 mice), TXNIP knockdown ( N = 5 mice), and RBMS3-TXNIP double knockdown ( N = 5 mice) in MDA-MB-231 cells. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. ANOVA was performed; bars indicate means and SEM. ( I ) Molecular mechanism of RBMS3 metastasis suppression through posttranscriptional regulatory action.

    Journal: Science Advances

    Article Title: Integrative analysis of mRNA stability regulation uncovers a metastasis-suppressive program in breast cancer

    doi: 10.1126/sciadv.aea9061

    Figure Lengend Snippet: ( A ) Schematic of the dual-guide CRISPR interference (CRISPRi) screen. ( B ) Analysis of CRISPRi screen comparing the abundance of cells expressing each guide between in vivo– and in vitro–grown cells with DESeq2. TXNIP was observed to be present at high levels in vivo and low levels in vitro indicating high metastasis but low proliferation. ( C ) Comparative analysis of TXNIP to RBMS3 expression in the METABRIC cohort . Shown is Pearson correlation with t statistic for nonzero correlation. ( D ) Analysis of disease-free survival in the METABRIC cohort relative to TXNIP expression. ( E ) Meta-analysis of relapse-free survival in smaller published cohorts relative to TXNIP expression. (D and E) The Mantel-Cox test was used to measure significance. Low TXNIP expression is indicative of poor prognosis for disease-free survival in patients with breast cancer. ( F ) The expression levels of TXNIP in 90 tumor samples at different stages of breast cancer was measured by qPCR; bars show means and SEM. ANOVA was performed. ( G ) The expression levels of TXNIP in RBMS3 knockdown MDA-MB-231 cells and control cells was measured by qPCR and compared using Mann-Whitney U test. ( H ) Lung colonization assays of control ( N = 5 mice), TXNIP knockdown ( N = 5 mice), and RBMS3-TXNIP double knockdown ( N = 5 mice) in MDA-MB-231 cells. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. ANOVA was performed; bars indicate means and SEM. ( I ) Molecular mechanism of RBMS3 metastasis suppression through posttranscriptional regulatory action.

    Article Snippet: We measured TXNIP expression using qPCR in 96 clinical samples across all stages of breast cancer, namely 5 normal epithelial, 23 stage I, 30 stage II, 29 stage III, and 9 stage IV metastatic biopsies (Origene, BCRT102, BCRT103), from which 90 samples yielded sufficient amount of cDNA.

    Techniques: CRISPR, Expressing, In Vivo, In Vitro, Knockdown, Control, MANN-WHITNEY, Luciferase, Labeling, Injection

    Figure 1. Expression of IL-13Rα2 in breast cancer tissue. (a) RT-PCR analysis of IL-13Rα2 mRNA expression in cDNA array samples (TissueScan™, Origene) derived from breast cancer (number of samples (n = 60)) and non-malignant (n = 7) tissue. Shown is the fold-change relative to mean expression in non-malignant samples (mean fold-change, cancer versus non-malignant; *** p ≤0.001). (b) Stratification analysis of IL-13Rα2 mRNA expression data comparing TNBC-type (n = 19) versus non-TNBC (n = 41) tumors (mean fold-change, TNBC versus non-TNBC, *** p ≤0.001). (c) Im- munohistochemical analysis of IL-13Rα2 protein expression in breast cancer tissue array samples (US Biomax, BR1009). Shown images (at 40× magnification) are representative examples of specific staining with an anti-IL-13Rα2 antibody (sc-134363) versus a mouse IgG2a isotype control: (left panel), non-malignant tissue; (middle panel) non-TNBC tumor (ER+, stage IIa); (right panel) TNBC tumor (stage IIb). (Right Figure), scoring analysis of anti-IL-13Rα2 reactivity in non-malignant (n = 9), non-TNBC (n = 8), and TNBC (n = 21) tissue array sections (mean score, TNBC versus non-malignant, *** p ≤0.001).

    Journal: Cancers

    Article Title: Targeting and Sensitization of Breast Cancer Cells to Killing with a Novel Interleukin-13 Receptor α2-Specific Hybrid Cytolytic Peptide.

    doi: 10.3390/cancers15102772

    Figure Lengend Snippet: Figure 1. Expression of IL-13Rα2 in breast cancer tissue. (a) RT-PCR analysis of IL-13Rα2 mRNA expression in cDNA array samples (TissueScan™, Origene) derived from breast cancer (number of samples (n = 60)) and non-malignant (n = 7) tissue. Shown is the fold-change relative to mean expression in non-malignant samples (mean fold-change, cancer versus non-malignant; *** p ≤0.001). (b) Stratification analysis of IL-13Rα2 mRNA expression data comparing TNBC-type (n = 19) versus non-TNBC (n = 41) tumors (mean fold-change, TNBC versus non-TNBC, *** p ≤0.001). (c) Im- munohistochemical analysis of IL-13Rα2 protein expression in breast cancer tissue array samples (US Biomax, BR1009). Shown images (at 40× magnification) are representative examples of specific staining with an anti-IL-13Rα2 antibody (sc-134363) versus a mouse IgG2a isotype control: (left panel), non-malignant tissue; (middle panel) non-TNBC tumor (ER+, stage IIa); (right panel) TNBC tumor (stage IIb). (Right Figure), scoring analysis of anti-IL-13Rα2 reactivity in non-malignant (n = 9), non-TNBC (n = 8), and TNBC (n = 21) tissue array sections (mean score, TNBC versus non-malignant, *** p ≤0.001).

    Article Snippet: To determine relative IL-13Rα2 mRNA expression in breast cancer tumors representing various stages of the disease, a similar RT-PCR methodology was applied to Origene TissueScanTM Breast Cancer cDNA Arrays (samples from CSRT104, BCRT103, and BCRT104; 60 tumor samples (n = 60) covering various disease stages with non-malignant (n = 7) breast tissue cDNA samples for comparison) [37].

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Staining, Control