Journal: BMC Genomics

Article Title: Cloning and characterization of the rat homologues of the Inhibitor of Apoptosis protein 1, 2, and 3 genes.

doi: 10.1186/1471-2164-3-5

Figure Lengend Snippet: ( A ) Northern blot analysis of rat iap mRNA expression in adult rat tissues. A rat multiple tissue northern blot (Clontech) containing 2 μg/lane poly(A) + RNA per lane was probed sequentially with [ 32 P] dCTP (Amersham) labeled, random primed (Amersham Rediprime) DNA probes derived from the coding regions of rat iap-1 , iap-2 , iap-3, and β-actin (control). Blots were hybridized overnight in 5 × SSPE/10 × Denhardt's solution/100 μg/ml salmon sperm DNA/ 50% formamide/ 2% SDS and then washed with 0.2 × SSC/ 0.1% SDS at 50°C. The position and sizes of the markers are indicated on the left. The tissues represented on the blot are as follows: ( 1 ) heart, ( 2 ) brain, ( 3 ) spleen, ( 4 ) lung, ( 5 ) liver, ( 6 ) skeletal muscle, ( 7 ) kidney, and ( 8 ) testis. ( B ) Northern blot analysis of the expression of iap -2 in testes. A rat multiple tissue Northern blot (Clontech; same as in (A)) was hybridized sequentially with [ 32 P] dCTP labeled DNA probes specific to iap -2 coding region, BIR domain and the RING zing finger. The blots were processed as in (A). The position of testis-specific transcript is indicated by an asterisk ( * ).

Article Snippet: A rat multiple tissue northern blot (Clontech) containing 2 μg/lane poly(A) + RNA per lane was probed sequentially with [ 32 P] dCTP (Amersham) labeled, random primed (Amersham Rediprime) DNA probes derived from the coding regions of rat iap-1 , iap-2 , and iap-3, or β-actin, respectively .

Techniques: Northern Blot, Expressing, Labeling, Random Primed, Derivative Assay

Journal: BMC Genomics

Article Title: Cloning and characterization of the rat homologues of the Inhibitor of Apoptosis protein 1, 2, and 3 genes.

doi: 10.1186/1471-2164-3-5

Figure Lengend Snippet: The relative expression levels of the rat iap genes in various tissues as detected by Northern or Western blot analysis.

Article Snippet: A rat multiple tissue northern blot (Clontech) containing 2 μg/lane poly(A) + RNA per lane was probed sequentially with [ 32 P] dCTP (Amersham) labeled, random primed (Amersham Rediprime) DNA probes derived from the coding regions of rat iap-1 , iap-2 , and iap-3, or β-actin, respectively .

Techniques: Expressing, Northern Blot, Western Blot

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