β actin c4  (Millipore)

Journal: Oncogene

Article Title: Integrin α1β1 expression is controlled by c-MYC in colorectal cancer cells

doi: 10.1038/onc.2015.231

Figure Lengend Snippet: MYC inhibition downregulates ITGA1 expression at the mRNA and protein levels in colorectal cancer cells. ( a ) T84, HT29 and SW480 cells were treated with the MYC inhibitor 10058-F4 used at 50 μ M in dimethyl sulfoxide (DMSO; MYCi) or with DMSO alone for the indicated times. Real-time quantitative PC R (qPCR) levels are indicated as fold changes of ITGA1 relative to the control and normalized to RPLP0 used as housekeeping gene. ( b ) Representative western blot and densitometric analyses of MYC and ITGA1 expression after treatment of T84, HT29 and SW480 cells with MYCi as in ( a ). β-Actin (ACTB) was used as loading control. All experiments were performed in triplicate and repeated three times. * P

Article Snippet: Membranes were then incubated overnight at 4 °C with the following primary antibodies: anti-human integrin α1/CD49a (1:2000, AF5676, R & D Systems, Minneapolis, MN, USA), anti-c-Myc (1:5000, Y69, ab32072, Abcam, Toronto, ON, Canada), anti-β1 (1:1000, BD Biosciences, Mississauga, ON, Canada) and anti-β-actin (1:80 000, C4, Millipore, Etobicoke, ON, Canada) as endogenous loading control.

Techniques: Inhibition, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal:

Article Title: Androgens act synergistically to enhance estrogen‐induced upregulation of human tissue kallikreins 10, 11, and 14 in breast cancer cells via a membrane bound androgen receptor), Androgens act synergistically to enhance estrogen‐induced upregulation of human tissue kallikreins 10, 11, and 14 in breast cancer cells via a membrane bound androgen receptor

doi: 10.1016/j.molonc.2008.01.001

Figure Lengend Snippet: Western blot analysis of hormone receptors. Cell lysates from BT474 were analyzed for changes in androgen (AR), estrogen (ER) and glucocorticoid (GR) and progesterone (PGR) receptor levels. β‐ACTIN is used as the loading control for this

Article Snippet: The antibodies used for Western blot analysis included, AR (N‐20), ERα (D‐12), and PGR (AB‐52), and β‐ACTIN (C4) from EMD Biosciences Inc., San Diego, CA and GR (Clone 41) from BD Biosciences, San Jose, CA.

Techniques: Western Blot

Journal: British Journal of Cancer

Article Title: SNPs at miR-155 binding sites of TYRP1 explain discrepancy between mRNA and protein and refine TYRP1 prognostic value in melanoma

doi: 10.1038/bjc.2015.194

Figure Lengend Snippet: Evaluation of TYRP1 regulation by miR-155 according to SNPs in melanoma cell lines. Measurement of TYRP1 mRNA and protein in ME1402 (CC/AA, ‘match' genotype) and MM031 (AA/CC, ‘mismatch' genotype) melanoma cell lines transfected with increasing concentrations of either pre-miR scramble (negative control) or pre-miR-155. ( A ) The miR-155 levels normalised to RNU44 (2 −ΔCT ) as function of pre-miR-155 used for transfection (basal level of miR-155 is 0.01 in MM031 and 0.03 in ME1402 line). ( B ) The TYRP1 mRNA expression normalised to RPS18 (2 −ΔCT ) as function of miR-155 measured in cells after transfection. ( C ) Representative western blots (WB) of TYRP1 protein ( β -actin as loading control) as function of the amount of pre-miR-155 used for transfection. ( D ) The TYRP1 protein WB bands normalised to β -actin as function of the amount of miR-155 measured in cells after transfection. All values are the mean of seven independent transfection experiments. No significant difference was found between data from pre-miR scramble transfected cells and nontransfected (control) cells. The TYRP1 mRNA and protein levels are normalized to values from control MM031 cells. Linear regression is used to assess the relationship between mRNA or protein and miR-155 levels.

Article Snippet: Immunodetection was performed using antibodies raised against TYRP1 (AB23, 1 : 1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and β -actin (C4, 1 : 5000, Millipore, Temecula, CA, USA).

Techniques: Transfection, Negative Control, Expressing, Western Blot