Journal: PloS one

Article Title: FABP7 and HMGCS2 are novel protein markers for apocrine differentiation categorizing apocrine carcinoma of the breast.

doi: 10.1371/journal.pone.0112024

Figure Lengend Snippet: Figure 7. Immunohistochemical analysis of molecular apocrine markers derived from trancriptomics data within the apocrine cyst and carcinoma dataset. Representative staining of FFPE of apocrine cysts and IACs with antibodies against SLC2A10 (Glucose transporter type 10, sections A and B) - cytoplasmic immunoreactivity; SLC7A8 (L-type amino acid transporter 2, sections C and D) - mainly cytoplasmic and membranous immunereactivity; MLPH (Melanophilin, sections E and F) - cytoplasmic and luminal membranous immunoreactivity; FOXA1 (Hepatocyte nuclear factor 3-alpha, sections G and H) – nuclear staining; XBP1 (X-box-binding protein 1, sections I and J) – mainly nuclear and cytoplasmic immunereactivity; BLVRA (Biliverdin reductase A, sections K and L) - cytoplasmic immunoreactivity which combined with rare nuclear positivity; RHOB (Rho-related GTP-binding protein RhoB, sections M and N) - cytoplasmic immunoreactivity; TSC22D3 (TSC22 domain family protein 3, sections O and P) - mainly nuclear and cytoplasmic immunereactivity; ABCA12 (ATP-binding cassette sub-family A member 12, sections Q and R) - cytoplasmic immunoreactivity and SIDT1 (SID1 transmembrane family member 1, sections I and G) - cytoplasmic immunoreactivity. Magnification: x10. Representative areas are shown in higher magnification (x20) and examples of positive staining are indicated by red arrows. doi:10.1371/journal.pone.0112024.g007

Article Snippet: Antibodies The polyclonal antibody raised against synthetic peptides of human FABP7 (HPA028825), BLVRA (HPA042865), ABCA12 (HPA043194), SIDT1 (HPA035862), MLPH (HPA014685) were purchased from Atlas Antibodies and were used in IHC at a dilution of 1:200, 1:100, 1:500, 1:50 and 1:100 respectively in accordance to the manufacturer instructions.

Techniques: Immunohistochemical staining, Derivative Assay, Staining, Binding Assay

Journal: PloS one

Article Title: FABP7 and HMGCS2 are novel protein markers for apocrine differentiation categorizing apocrine carcinoma of the breast.

doi: 10.1371/journal.pone.0112024

Figure Lengend Snippet: Figure 8. Protein expression profile of transcriptome-derived apocrine markers among breast cancer subtypes. The diagram presents immunohistochemical profiles of four breast tumor subtypes, TNBC, Luminal A Luminal B and HER2, reacted with antibodies against ten transcriptome-derived apocrine markers: XBP1, TSC22D3, ABCA12, SIDT1, FOXA1, RHOB, BLVRA, SLC2A10, SLC7A8 and MLPH. The rows indicate the expression of particular protein in every core in breast cancer TMA (BRC1501, 1502 and 1503; Pantomics INC, USA): red box – positive staining, white box – negative staining grey box – parameter was not determined. Corresponding frequencies of positives are shown on the right side of the diagram. Samples were considered as positive if 10% or more of the cells showed a clear positive staining with the antibodies. IDC = invasive ductal carcinoma; TNBC = triple negative breast cancer. Tumors have been stratified as specified in Materials and Methods. doi:10.1371/journal.pone.0112024.g008

Article Snippet: Antibodies The polyclonal antibody raised against synthetic peptides of human FABP7 (HPA028825), BLVRA (HPA042865), ABCA12 (HPA043194), SIDT1 (HPA035862), MLPH (HPA014685) were purchased from Atlas Antibodies and were used in IHC at a dilution of 1:200, 1:100, 1:500, 1:50 and 1:100 respectively in accordance to the manufacturer instructions.

Techniques: Expressing, Derivative Assay, Immunohistochemical staining, Staining, Negative Staining

Journal: PloS one

Article Title: FABP7 and HMGCS2 are novel protein markers for apocrine differentiation categorizing apocrine carcinoma of the breast.

doi: 10.1371/journal.pone.0112024

Figure Lengend Snippet: Figure 9. MLPH is expressed by non-malignant apocrine cells but is lost in IACs. Representative images of FFPE sections immunostained with antibody against MLPH. (A) normal ducts, (B) benign apocrine cysts (mainly luminal membranous immunoreactivity), (C) sclerosing adenosis with apocrine differentiation, (D) IAC showing positive immunostaining in pseudo-glands structures (cytoplasmic and luminal membranous immunoreactivity) and (E) IAC with negative immunostaining. Magnification: x20. doi:10.1371/journal.pone.0112024.g009

Article Snippet: Antibodies The polyclonal antibody raised against synthetic peptides of human FABP7 (HPA028825), BLVRA (HPA042865), ABCA12 (HPA043194), SIDT1 (HPA035862), MLPH (HPA014685) were purchased from Atlas Antibodies and were used in IHC at a dilution of 1:200, 1:100, 1:500, 1:50 and 1:100 respectively in accordance to the manufacturer instructions.

Techniques: Immunostaining

Journal: Molecular carcinogenesis

Article Title: Single nucleotide polymorphism rs13426236 contributes to an increased prostate cancer risk via regulating MLPH splicing variant 4

doi: 10.1002/mc.23127

Figure Lengend Snippet: A. RT-qPCR shows a significant increase of MLPH RNA transcripts after 48h transfection. B. Western blot analysis shows a significant increase of MLPH protein after 48h transfection. Asterisk (*) indicates that a single beta-actin band was split in sample DU145 control due to broken gel during membrane transfer. C. Cell viability test shows significantly higher live cell population in transcript V4-transfected cells than vector control. However, transcript V1-transfected cells did not show significant increase in live cell population when compared to vector control.

Article Snippet: The membrane was incubated with MLPH antibody (LSBio, Seattle, WA, USA) overnight after blocked in 3% bovine serum albumin (BSA) in TBST (Tris-buffered saline, 0.1% Tween 20).

Techniques: Quantitative RT-PCR, Transfection, Western Blot, Plasmid Preparation