polyclonal anti-dctn3 primary antibody  (Millipore)

Journal: Cancer Medicine

Article Title: Recurring DNA copy number gain at chromosome 9p13 plays a role in the activation of multiple candidate oncogenes in progressing oral premalignant lesions

doi: 10.1002/cam4.307

Figure Lengend Snippet: Candidate gene silencing contributes to decreased cell proliferation in SCC-9 cells (an oral cancer cell line harboring the recurrent 9p13 amplicon). (A) presents VCP, (B) DCTN3, (C) STOML2. Each panel contains three experiments performed for each gene, presenting images of, knockdown efficiencies (qRT-PCR) for five different shRNAs against the candidate gene, protein expression of two best knockdowns (Western blots), and MTT proliferation results over a 5-day period of the two confirmed knockdowns. (D) shRNA stable knockdown of each candidate gene in SCC-9 cells decreases colony formation in soft agar relative to control SCC-9 cells. The two most effective shRNAs for each candidate gene are shown. Triplicate experiments were performed for each line. The mean number of colonies and standard deviations are plotted.

Article Snippet: Membrane blocking was performed in 5% w/v nonfat dry milk, 1× TBS, and 0.1% Tween-20 at room temperature with gentle shaking for 1 h for polyclonal anti-DCTN3 primary antibody (Sigma-Aldrich).

Techniques: Amplification, Quantitative RT-PCR, Expressing, Western Blot, shRNA

Journal: Cancer Medicine

Article Title: Recurring DNA copy number gain at chromosome 9p13 plays a role in the activation of multiple candidate oncogenes in progressing oral premalignant lesions

doi: 10.1002/cam4.307

Figure Lengend Snippet: Stable overexpression of STOML2 (A), VCP (B), and DCTN3 (C) increases cell growth in Cal-27 cells (an oral cancer cell line not exhibiting the 9p13 amplicon). MTT proliferation results over a 5-day period are shown. Each panel for each gene depicts (1) an MTT cell proliferation plot for both control and test lines, (2) qRT-PCR results demonstrating increased expression of a given candidate gene in test line, and (3) Western blots confirming concordant protein overexpression of test line. (D) Stable overexpression of candidate gene in Cal-27 cells increases colony growth in soft agar relative to control Cal-27 cells. Experiments were performed in triplicate. Mean number of colonies and standard deviations are plotted.

Article Snippet: Membrane blocking was performed in 5% w/v nonfat dry milk, 1× TBS, and 0.1% Tween-20 at room temperature with gentle shaking for 1 h for polyclonal anti-DCTN3 primary antibody (Sigma-Aldrich).

Techniques: Over Expression, Amplification, MTT Cell Proliferation, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cancer Medicine

Article Title: Recurring DNA copy number gain at chromosome 9p13 plays a role in the activation of multiple candidate oncogenes in progressing oral premalignant lesions

doi: 10.1002/cam4.307

Figure Lengend Snippet: Stable overexpression of STOML2 (A), VCP (B), and DCTN3 (C) increases cell growth in the dysplasia cells line, POE9n-TERT (an oral dysplasia cell line with no 9p13 amplicon). MTT proliferation results over a 5-day period are shown. Each panel for each gene depicts (1) an MTT cell proliferation plot for both control and test lines, (2) qRT-PCR results demonstrating increased expression of a given candidate gene in test line, and (3) Western blots confirming increased amount of protein of test line.

Article Snippet: Membrane blocking was performed in 5% w/v nonfat dry milk, 1× TBS, and 0.1% Tween-20 at room temperature with gentle shaking for 1 h for polyclonal anti-DCTN3 primary antibody (Sigma-Aldrich).

Techniques: Over Expression, Amplification, MTT Cell Proliferation, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cancer Medicine

Article Title: Recurring DNA copy number gain at chromosome 9p13 plays a role in the activation of multiple candidate oncogenes in progressing oral premalignant lesions

doi: 10.1002/cam4.307

Figure Lengend Snippet: DNA gain at 9p13 and corresponding increased mRNA expressions in gene candidates detected in biopsies from a single patient. Clinical, pathological, and molecular characterization of different tissues obtained from an oral cancer disease field in a single patient. (A) to (D) are photomicrographs of hematoxylin and eosin stained slides of varying histology: (A) normal, (B) mild dysplasia, (C) carcinoma in situ (CIS), and (D) invasive squamous cell carcinoma (SCC) (original magnification, 200×). (E) White light image of the oral cancer at right side of tongue. Location of biopsies and its corresponding histology (A–D) are indicated. (F) Fluorescence visualization (FV) image of the same lesion, with a broad area of dark brown change in FV loss. (G) Alignment of genomic alteration data at chromosome 9p indicates 9p13 amplification in the CIS and SCC cells obtained from this single patient. The red vertical lines indicate log2 signal intensity ratios of +1 and +0.5 and the green lines indicate ratios of −1 and −0.5. Each black dot represents the log2 signal intensity ratio of an individual clone mapping to the array. (H) Increased fold-change mRNA expressions (>twofold) are found for STOML2, VCP, and DCTN3 as determined by gene expression microarray. No standard error is indicated for STOML2 as only a single probe maps to this gene.

Article Snippet: Membrane blocking was performed in 5% w/v nonfat dry milk, 1× TBS, and 0.1% Tween-20 at room temperature with gentle shaking for 1 h for polyclonal anti-DCTN3 primary antibody (Sigma-Aldrich).

Techniques: Staining, In Situ, Fluorescence, Amplification, Expressing, Microarray