Journal: Cancer Science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Fhl1 and Sdpr expression in normal epithelium and tumors from human breast, kidney, and prostate. Immunohistochemical staining was performed on normal and tumor human tissue. Fhl1 and Sdpr staining was evident in normal epithelial cells of the breast, kidney, and prostate. In contrast, both Fhl1 and Sdpr expression were suppressed in infiltrating mammary duct carcinoma, renal cell carcinoma, and prostate adenocarcinoma. (bar = 60 microns).

Article Snippet: Affinity‐purified (Pierce MicroLink Peptide Coupling Kit, Cat No: 20485) rabbit antiserum targeted against Fhl1 (Proteintech Group, S1009) or Sdpr (Proteintech Group, 90124) was applied to the sections and incubated at 4°C overnight in IHC blocking buffer.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Cancer Science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Fhl1 and Sdpr expression in human breast, kidney, and prostate tumors

Article Snippet: Affinity‐purified (Pierce MicroLink Peptide Coupling Kit, Cat No: 20485) rabbit antiserum targeted against Fhl1 (Proteintech Group, S1009) or Sdpr (Proteintech Group, 90124) was applied to the sections and incubated at 4°C overnight in IHC blocking buffer.

Techniques: Expressing

Journal: Cancer Science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Selection of genes affected by Fhl1. Microarray analysis was performed to identify probe sets representing genes with mean fold change differences of 2 or more with P < 0.05 by t‐test. (a) Fhl1 transfected Src transformed cells were compared with control transfectants. Of approximately 45 000 probe sets examined, genes represented by 459 were induced by Fhl1, whereas genes represented by 189 were suppressed. (b) Non‐transformed cells were compared with Src transformed cells to identify genes affected by Fhl1 that were correspondingly regulated by Src. Genes represented by 53 probe sets were induced by Fhl1 and suppressed by Src, whereas 48 were suppressed by Fhl1 and induced by Src. (c) Src‐transformed homozygous null Cas knockout cells transfected with Cas were compared with control transfectants to identify genes correspondingly regulated by Fhl1, Scr, and Cas. Genes represented by four probe sets were induced by Fhl1 and suppressed by Src and Cas, whereas genes represented by two probe sets were suppressed by Fhl1 and induced by Src and Cas.

Article Snippet: Affinity‐purified (Pierce MicroLink Peptide Coupling Kit, Cat No: 20485) rabbit antiserum targeted against Fhl1 (Proteintech Group, S1009) or Sdpr (Proteintech Group, 90124) was applied to the sections and incubated at 4°C overnight in IHC blocking buffer.

Techniques: Selection, Microarray, Transfection, Transformation Assay, Control, Knock-Out

Journal: Cancer Science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Effects of Src and mitogen‐activated protein kinase (MAPK) activity on Fhl1 and Sdpr expression. (a) An equal amount of total protein (10 µg/lane) from non‐transformed and Src‐transformed NIH 3T3 cells expressing a Central Dogma (CD)‐tagged Sdpr–GFP fusion protein was analyzed by Western blotting with antiserum specific for green fluorescence protein (GFP), Sdpr, mitogen‐activated protein kinase kinase (MEK), MAPK, active MAPK (p‐MAPK), active Src (p‐Src), and β‐actin as indicated. (b) An equal amount of protein (100 µg) was immunoprecipitated from non‐transformed and Src‐transformed NIH 3T3 cells expressing a CD‐tagged Sdpr–GFP fusion protein, and then examined by Western blot analysis with GFP or Sdpr antiserum. (c) An equal amount of total protein (10 µg/lane) from non‐transformed and Src‐transformed embryonic mouse brain cells was analyzed by Western blotting with antiserum specific for Sdpr, Fhl1, MAPK, active MAPK (p‐MAPK), active Src (p‐Src), and β‐actin as indicated. (d) An equal amount of total protein (10 µg/lane) from non‐transformed and Src transformed embryonic mouse fibroblasts with or without Cas was analyzed by Western blotting with antiserum specific for Fhl1, MAPK, active MAPK (p‐MAPK), Cas, active Src (p‐Src), and β‐actin as indicated. Src‐transformed cells were also treated with the MEK blocker PD98059 to determine if Src requires MAPK to suppress Fhl1 and Sdpr expression. In all cases, more Fhl1 and Sdpr protein was present in non‐transformed cells than Src‐transformed cells independent of MAPK activity.

Article Snippet: Affinity‐purified (Pierce MicroLink Peptide Coupling Kit, Cat No: 20485) rabbit antiserum targeted against Fhl1 (Proteintech Group, S1009) or Sdpr (Proteintech Group, 90124) was applied to the sections and incubated at 4°C overnight in IHC blocking buffer.

Techniques: Activity Assay, Expressing, Transformation Assay, Western Blot, Fluorescence, Immunoprecipitation

Journal: Cancer Science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Correlation of Fhl1 and Sdpr expression with Gleason stage of prostate tumors

Article Snippet: Affinity‐purified (Pierce MicroLink Peptide Coupling Kit, Cat No: 20485) rabbit antiserum targeted against Fhl1 (Proteintech Group, S1009) or Sdpr (Proteintech Group, 90124) was applied to the sections and incubated at 4°C overnight in IHC blocking buffer.

Techniques: Expressing

Journal: Cancer Science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Fhl1 gene methylation in Src transformed cells. The transcriptional start site of Fhl1 (+1) lies in a CpG island of several hundred residues. PCR products from bisulfate‐treated DNA from non‐transformed and Src‐transformed cells were subcloned and sequenced. Data are shown as the percentage of each CpG sequence that was methylated (n = 5). More methylation of most of these sites was found in Src‐transformed cells than in non‐transformed cells.

Article Snippet: Affinity‐purified (Pierce MicroLink Peptide Coupling Kit, Cat No: 20485) rabbit antiserum targeted against Fhl1 (Proteintech Group, S1009) or Sdpr (Proteintech Group, 90124) was applied to the sections and incubated at 4°C overnight in IHC blocking buffer.

Techniques: Methylation, Transformation Assay, Sequencing

Journal: Cancer Science

Article Title: Coordinate suppression of Sdpr and Fhl1 expression in tumors of the breast, kidney, and prostate

doi: 10.1111/j.1349-7006.2008.00816.x

Figure Lengend Snippet: Genes regulated by Fhl1

Article Snippet: Affinity‐purified (Pierce MicroLink Peptide Coupling Kit, Cat No: 20485) rabbit antiserum targeted against Fhl1 (Proteintech Group, S1009) or Sdpr (Proteintech Group, 90124) was applied to the sections and incubated at 4°C overnight in IHC blocking buffer.

Techniques: