Journal: Scientific Reports

Article Title: A tool kit for rapid cloning and expression of recombinant antibodies

doi: 10.1038/srep05885

Figure Lengend Snippet: (A–B) Flow cytometric histograms showing fluorescent intensity of cells incubated with purified IgG/IgE antibodies plus secondary FITC-labelled goat anti-human IgG/IgE (open) and cells with secondary antibody only (filled) against the number of cells. (A) Flow cytometric assessment of anti-CSPG4 IgG 1 /IgE antibodies shows specific binding to native CSPG4 antigen present on the cell surface of A375 melanoma cells and no binding above background to primary melanocytes. (B) Fc regions of anti-CSPG4 IgG 1 and 102.1F10 IgG 4 isotypes demonstrate effector-binding to U937 monocytic cell line, expressing human Fcγ receptors. The IgE antibody isotypes also bind similarly to RBL SX38 mast cells, expressing human FcεR1 receptor. (C) Immunofluorescence staining of A375 cells confirms specific binding of anti-CSPG4 IgG 1 /IgE and no background binding with isotype control hapten specific NIP-IgG 1 /IgE detected by goat anti-human IgG/IgE-FITC. (D) Grass pollen allergen specificity is confirmed by anti-human sandwich ELISA, showing specific binding of recombinant 102.1F10 IgG 4 /IgE and original patient serum to the ELISA plate-bound Phl p 7 allergen and no binding above background with unspecific human myeloma IgG 4 and MOv18 IgE isotype controls.

Article Snippet: Primary human epidermal melanocytes (PCS-200-012, ATCC) were grownin Dermal Cell Basal Medium (PCS-200-030, ATCC) and supplemented with the Melanocyte Growth Kit (PCS-200-041, ATCC).

Techniques: Incubation, Purification, Binding Assay, Expressing, Immunofluorescence, Staining, Control, Sandwich ELISA, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: PloS one

Article Title: CCHCR1 is up-regulated in skin cancer and associated with EGFR expression.

doi: 10.1371/journal.pone.0006030

Figure Lengend Snippet: Figure 7. CCHCR1 mRNA expression in normal keratinocytes and SCC cell lines correlates with Ki67 expression. A) Ki67, EGFR, cyclin- D1 and CCHCR1 gene expression profile of five normal epidermal KC and eight cutaneous SCC cell lines (heatmap). Signal values of the probe sets were compared to the mean signal values of each probe set in KCs. The colouring is based on the log2 values of the change in the signal values. The up-regulated genes are shown in red and down-regulated genes are shown in green. B) Correlation between CCHCR1 and Ki67 probe sets was calculated between the signal values of one CCHCR1 and one Ki67 probe set in the HG-U133 Plus 3.0 array. Pearson’s correlation coefficient R = 0.88. C) CCHCR1 and D) Ki67 mRNA expression levels in the normal KCs and cutaneous SCC cell lines as measured by qRT-PCR (TaqMan). Expression levels of CCHCR1 and Ki67 in the normal KCs and SCC cell lines were analyzed by qRT-PCR and corrected for the b-actin mRNA levels in the same samples. E) Correlation between CCHCR1 and Ki67 probe sets was calculated between the signal values of CCHCR1 and Ki67 mRNA levels. Pearson’s correlation coefficient R = 0.46. doi:10.1371/journal.pone.0006030.g007

Article Snippet: Normal human epidermal KC cell lines (n = 5) were established from skin samples, and one NHEK cell lines was purchased from PromoCell (Heidelberg, Germany).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR

Journal: Human Mutation

Article Title: A Macrophage/Monocyte‐Related Four‐Gene Signature for Prognostic Assessment of Uveal Melanoma: BTBD6 , C2CD4B , CCL24 , and S100A4

doi: 10.1155/humu/4978880

Figure Lengend Snippet: Role of four prognostically relevant genes. (A) Quantitative reverse transcription PCR for the calculated levels of four prognostically relevant genes in human epidermal melanocytes. (B) Validation of the knockdown efficiency of C2CD4B. (C, D) Scratch and Transwell assays exploring the migration and invasion of A375 cells in vitro. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, p > 0.05.

Article Snippet: Melanoma cell line A375 (CRL‐1619) and human epidermal melanocytes (PCS‐200‐013) were purchased from American Type Culture Collection (ATCC, Manassas, Virginia) and cultured as follows.

Techniques: Reverse Transcription, Biomarker Discovery, Knockdown, Migration, In Vitro