Journal: Breast Cancer Research

Article Title: Involvement of maternal embryonic leucine zipper kinase (MELK) in mammary carcinogenesis through interaction with Bcl-G, a pro-apoptotic member of the Bcl-2 family

doi: 10.1186/bcr1650

Figure Lengend Snippet: Expression and distribution of MELK in human normal tissues and breast cancer cell lines. (a) Expression of MELK in 12 breast cancer specimens (case number; 42, 102, 247, 252, 302, 473, 478, 502, 552, 646, 769 and 779) by semi-quantitative RT-PCR. GAPDH served as a quantitative internal control. (b) Multiple tissue Northern blot analysis demonstrated that an approximately 2.7 kb MELK transcript was detected in the testis, thymus and small intestine. PBL, peripheral blood leukocytes. (c) Breast cancer cell line Northern blot analysis revealed that approximately 2.4 to 2.7 kb MELK variants were specifically expressed in breast cancer cell lines, but not in normal vital organs. (d) Schematic representation of three variant transcripts identified by cDNA library screening (see Materials and methods). White boxes indicate a coding region and black boxes indicate a non-coding region. Black and grey triangles indicate initiation codons, and white triangles indicate stop codons. Exon numbers are shown above each box. (e) In vitro translation assay of each variant isolated from cDNA library screening. The number within parentheses represents the predicted molecular weight (kDa) of each variant protein. (f) Expression of MELK proteins in eight breast cancer cell lines as well as human mammary epithelial cells (HMECs) shown by western blot analysis with an anti-MELK antibody. β-Actin served as a control. (g) Schematic representation of the V1, V2 and V3 forms of MELK. The shaded boxes indicate the catalytic domain (amino acids 11 to 263 of the V1 protein). The KA1 domain is the kinase-associated domain in the carboxy-terminal region.

Article Snippet: Breast cancer Northern blots and human multiple-tissue Northern blots (Takara Clontech) were hybridized with [α 32 P]-dCTP-labeled PCR products of MELK cDNA prepared by RT-PCR (see below).

Techniques: Expressing, Quantitative RT-PCR, Northern Blot, Variant Assay, cDNA Library Assay, In Vitro, Isolation, Molecular Weight, Western Blot

Journal: PLoS ONE

Article Title: Identification of MAMDC1 as a Candidate Susceptibility Gene for Systemic Lupus Erythematosus (SLE)

doi: 10.1371/journal.pone.0008037

Figure Lengend Snippet: Northern blot analysis of MAMDC1 mRNA expression in different human tissues, showing several expressed splice variants. MAMDC1 mRNA transcript corresponding to the full-length isoform is indicated by an arrowhead on the right side. A β-Actin cDNA control probe was used for normalization (bottom).

Article Snippet: Fifty nano grams (ng) of purified cDNA were then labelled with P 32 -dCTP (GE Healthcare, Buckinghamshire, UK) by random priming and was used to probe two human multiple tissue polyA+ RNA Northern blots (HB2010 and HB2011, OriGene Technologies) according to manufacturer's instructions.

Techniques: Northern Blot, Expressing

Journal: PLoS ONE

Article Title: Identification of MAMDC1 as a Candidate Susceptibility Gene for Systemic Lupus Erythematosus (SLE)

doi: 10.1371/journal.pone.0008037

Figure Lengend Snippet: THP-1 cells were treated for 24 h with LPS, TGF-β1, IL-1β, IFN-γ, TNF-α, or a combination of TNF-α and IFN-γ, and MAMDC1 mRNA expression was quantified by Real-Time PCR in triplicate experiments. The results are reported as fold changes relative to THP-1 cells grown in the absence of stimulation (control), with the smallest observation, lower quartile, median, upper quartile, and largest observation shown for each sample.

Article Snippet: Fifty nano grams (ng) of purified cDNA were then labelled with P 32 -dCTP (GE Healthcare, Buckinghamshire, UK) by random priming and was used to probe two human multiple tissue polyA+ RNA Northern blots (HB2010 and HB2011, OriGene Technologies) according to manufacturer's instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Upregulation of Nuclear Factor-Related Kappa B Suggests a Disorder of Transcriptional Regulation in Minimal Change Nephrotic Syndrome

doi: 10.1371/journal.pone.0030523

Figure Lengend Snippet: A, Western blot of total lysates using anti-NFRKB reveals a double band at the expected size. The specificity was attested by the loss of the binding upon preincubation of antibody with synthetic peptides corresponding to positions 543–557 and 1091–1106 within the primary structure of NFRKB. These data are representative of three independent experiments. B, Northern blot analysis of NFRKB. Multiple tissue-Northern blot (Clontech Laboratories, Inc.) was hybridized with a full-length-probe and exposed to Phosphoimager Storm for 24 h. This experiment was repeated once.

Article Snippet: Human multiple-tissue Northern blot (CLONTECH Laboratories, Inc.) was hybridized with a cDNA probe corresponding to the full-length coding sequence.

Techniques: Western Blot, Binding Assay, Northern Blot