Journal: Breast Cancer Research : BCR

Article Title: Regulator of G-protein signalling 2 mRNA is differentially expressed in mammary epithelial subpopulations and over-expressed in the majority of breast cancers

doi: 10.1186/bcr1834

Figure Lengend Snippet: Multiple staining protocols for flow cytometric analysis of α-Sca-1/33A10/α-CD45 and α-Sca-1/33A10/α-CD45/α-CD24 stained cells.

Article Snippet: After a further 30 minutes at room temperature the samples were analyzed on a Becton Dickenson LSRII flow cytometric analyser (Becton Dickenson, Oxford, UK).

Techniques: Staining, Control

Journal: Breast Cancer Research : BCR

Article Title: Regulator of G-protein signalling 2 mRNA is differentially expressed in mammary epithelial subpopulations and over-expressed in the majority of breast cancers

doi: 10.1186/bcr1834

Figure Lengend Snippet: Isolation and characterisation of mammary epithelial cell subpopulations. (a) Flow cytometric staining profiles (dead and CD45 + cells excluded) of anti-Sca-1 and 33A10 stained, freshly isolated mouse mammary cell preparations together with nonspecific IgG-stained control. (b) Graphical space representation of principal component analysis of mammary fibroblasts, Sca-1 + 33A10 Low/- , Sca-1 - 33A10 High and Sca-1 - 33A10 Low/- cells. (c) Mean fold differences ± 95% confidence limits in RNA abundance measured by quantitative real-time PCR for estrogen receptor ( Esr1 ) and prolactin receptor ( Prlr ) transcripts in Sca-1 + 33A10 Low/- ( n = 5 samples) and Sca-1 - 33A10 High ( n = 3 samples) mammary subpopulations compared with bulk mammary cell preparations depleted for CD45 + cells (comparator; n = 3 samples). The dotted lines indicate the 95% confidence limits of the comparator sample. All samples show a significant difference to the comparator (** P < 0.01) [35].

Article Snippet: After a further 30 minutes at room temperature the samples were analyzed on a Becton Dickenson LSRII flow cytometric analyser (Becton Dickenson, Oxford, UK).

Techniques: Isolation, Staining, Control, Real-time Polymerase Chain Reaction

Journal: Breast Cancer Research : BCR

Article Title: Regulator of G-protein signalling 2 mRNA is differentially expressed in mammary epithelial subpopulations and over-expressed in the majority of breast cancers

doi: 10.1186/bcr1834

Figure Lengend Snippet: Rgs2 is highly expressed in CD24 Low Sca-1 - 33A10 - mouse mammary basal/myoepithelial and human breast myoepithelial cells. (a) Flow cytometric staining profile of mouse mammary cell preparations stained with anti-Sca-1 and anti-CD24 antibodies together with either a nonspecific rat IgG and anti-rat-FITC or 33A10 and anti-rat FITC. The nonspecific IgG and 33A10 staining profiles of the CD24 Low Sca-1 - basal/myoepithelial cells (33A10 - ), CD24 High Sca-1 - (Esr1 - ) luminal cells (33A10 High ) and CD24 High Sca-1 + (Esr1 + ) luminal cells (33A10 Low ) [14] are indicated. (b) Mean fold differences ± 95% confidence limits in RNA abundance for the Rgs2 gene in CD24 Low Sca-1 - basal/myoepithelial, CD24 High Sca-1 - (Esr1 - ) luminal epithelial and CD24 High Sca-1 + (Esr1 + ) luminal epithelial mouse mammary cells ( n = 3 for all samples) [14]. The dotted lines indicate the 95% confidence limits of the comparator sample. All samples show a significant difference to the comparator (** P < 0.01). The basal myoepithelial cells also have a significantly higher level of Rgs2 expression than either of the two luminal populations. (c) Mean fold differences ± 95% confidence limits in expression levels for the RGS2 gene in myoepithelial and luminal epithelial human breast cells compared with human breast fibroblasts (comparator). See Materials and methods for details of the samples. The dotted lines indicate the 95% confidence limits of the comparator sample. Both samples show a significant difference to the comparator (** P < 0.01) and the myoepithelial cells have a significantly higher level of RGS2 expression than the luminal cells.

Article Snippet: After a further 30 minutes at room temperature the samples were analyzed on a Becton Dickenson LSRII flow cytometric analyser (Becton Dickenson, Oxford, UK).

Techniques: Staining, Expressing