Journal:

Article Title: Differential Regulation of Human Thymosin Beta 15 Isoforms by Transforming Growth Factor Beta 1

doi: 10.1002/gcc.20659

Figure Lengend Snippet: Primer and siRNA Sequences

Article Snippet: The Cancer Survey I qPCR tissue array (OriGene Technologies, Rockville, MA) was used to assess the expression of TMSB15A and TMSB15B in various tissues.

Techniques: Sequencing

Journal:

Article Title: Differential Regulation of Human Thymosin Beta 15 Isoforms by Transforming Growth Factor Beta 1

doi: 10.1002/gcc.20659

Figure Lengend Snippet: Alignment of TMSB15A and TMSB15B isoform mRNAs. The coding sequence is boxed, with the amino acid sequence shown above. Nucleotide sequence differences are indicated in bold and exon splice sites are marked with arrowheads. Primer locations are underlined.

Article Snippet: The Cancer Survey I qPCR tissue array (OriGene Technologies, Rockville, MA) was used to assess the expression of TMSB15A and TMSB15B in various tissues.

Techniques: Sequencing

Journal:

Article Title: Differential Regulation of Human Thymosin Beta 15 Isoforms by Transforming Growth Factor Beta 1

doi: 10.1002/gcc.20659

Figure Lengend Snippet: TMSB15A and TMSB15B are expressed at different levels in different human tumor cells and normal tissues. (A) Specific primers were used to amplify TMSB15A or TMSB15B in various tumor cells. HCT116, DU145, LNCaP-LN3 and LNCaP cells expressed more TMSB15A (labeled A) than TMSB15B (labeled B) (left panel). MCF-7, SKOV-3, HT1080 and PC-3MLN4 cells expressed approximately equivalent levels of each (center panel), while PC-3, MDA-MB-231, NCI-H322 and Caco-2 cells expressed more TMSB15B than TMSB15A (right panel). B2M amplified as control. (B) Left panel; quantitative PCR analysis of TMSB15A (solid bars) and TMSB15B (striped bars) in prostate showed TMSB15A was the main isoform expressed in either normal (N) or cancer (C) tissue. Data shown as fold expression relative to TMSB15B C(t) value for each prostate tissue sample. Right panel; quantitative PCR analysis in colon showed TMSB15B was the major isoform expressed in either normal or tumor tissue. Data shown as fold expression relative to TMSB15A C(t) value for each colon tissue sample.

Article Snippet: The Cancer Survey I qPCR tissue array (OriGene Technologies, Rockville, MA) was used to assess the expression of TMSB15A and TMSB15B in various tissues.

Techniques: Labeling, Amplification, Real-time Polymerase Chain Reaction, Expressing

Journal:

Article Title: Differential Regulation of Human Thymosin Beta 15 Isoforms by Transforming Growth Factor Beta 1

doi: 10.1002/gcc.20659

Figure Lengend Snippet: TMSB15A and TMSB15B RNA expression are differentially regulated by TGFB1. MCF-7 breast cancer cells were treated with 10 ng/ml TGFB1 or control conditions for 24 or 48 hrs. qPCR was used to assess RNA expression of TMSB15A and TMSB15B. TMSB15A levels (black bars, marked ‘A’) were significantly inhibited by TGFB1, while no effect was observed on TMSB15B (striped bars, marked ‘B’), * P < 0.05, ** P < 0.01. Fold change relative to RPS9 housekeeping gene expression and error bars indicate standard deviation of replicate samples.

Article Snippet: The Cancer Survey I qPCR tissue array (OriGene Technologies, Rockville, MA) was used to assess the expression of TMSB15A and TMSB15B in various tissues.

Techniques: RNA Expression, Expressing, Standard Deviation

Journal:

Article Title: Differential Regulation of Human Thymosin Beta 15 Isoforms by Transforming Growth Factor Beta 1

doi: 10.1002/gcc.20659

Figure Lengend Snippet: RNA interference demonstrates function of TMSB15B in cell migration to EGF. (A) PC-3MLN4 cells were treated with 3 individual control siRNAs, 2 TMSB15A siRNAs (#9 and #10) or 3 TMSB15B siRNAs (#2, #3 and #4). RT-PCR analysis using primers to TMSB15A (top panel), TMSB15B (center panel), or B2M housekeeping gene (lower panel) demonstrated isoform specificity of siRNA duplexes. (B) siRNA inhibition of TMSB15B significantly reduced PC-3 cell migration to EGF in the modified Boyden chamber assay. Data shown as mean migrated cells/low power field with error bars indicating standard error of the mean. Empty bar; untransfected cells, Horizontal striped bars; control siRNAs (thin stripe control siRNA#1, thick stripe control siRNA#3), Solid bars; TMSB15A siRNAs (solid black; TMSB15A siRNA#9, solid grey; TMSB15A siRNA#9), Striped bars; TMSB15B siRNAs (thin upward stripe; TMSB15B siRNA#2, thick downward stripe; TMSB15B siRNA#3, thick upward stripe; TMSB15B siRNA#4). * P < 0.01. (C) Parallel transfections were performed in PC-3 cells for RNA collection. RT-PCR showed decreased TMSB15B expression in all TMSB15B siRNA-treated cells. PC-3MLN4 was used as a positive control for PCR reactions (+). B2M; beta-2-microglobulin.

Article Snippet: The Cancer Survey I qPCR tissue array (OriGene Technologies, Rockville, MA) was used to assess the expression of TMSB15A and TMSB15B in various tissues.

Techniques: Migration, Reverse Transcription Polymerase Chain Reaction, Inhibition, Modification, Boyden Chamber Assay, Transfection, Expressing, Positive Control