Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: α-Klotho Expression in Human Tissues

doi: 10.1210/jc.2015-1800

Figure Lengend Snippet: α-Klotho isoforms, sequence, and Western blot. A, Structure of the two isoforms of α-Klotho. Isoform 1 represents the full-length protein and contains a signal sequence domain (SS), two homologous domains (KL1, KL2), a short transmembrane domain (TM), and a short cytoplasmic tail. Shown is the site of the epitope for the antibody used in our experiments: AA 800 to 900, KL2. This epitope is absent from Isoform 2, a soluble, secreted protein that arises from alternative RNA splicing and contains only AA 1–549, and where the terminal 15 residues are replaced by the sequence shown. B, The full-length α-Klotho protein sequence of 1012 AA is shown, with KL1 and KL2 shown in red and green respectively, and TM highlighted (black). The peptides giving rise to the PRM signature are also shown (bold typeface; common to isoforms 1 and 2, red; exclusive to full-length α-Klotho, isoform 1, blue). C and D, Western blot analysis of cell lysates (C) and tissues (D) supports the presence of the full-length α-Klotho. Full-length rh α-Klotho protein (rh-α-Klotho). EC, epithelial cells.

Article Snippet: Cell lysates were obtained from commercially available primary cell cultures (ScienCell Research Laboratories), including: Human Epidermal Keratinocyte Lysate-adult (product code HEKL-a, catalog no. 2116), Human Prostate Epithelial Cell Lysate (product code HPrEpiCL, catalog no. 4416), Human Mammary Epithelial Cell Lysate (product code HMEpiCL, catalog no. 7616), Human Renal Proximal Tubular Epithelial Cell Lysate (product code HRPTEpiCL, catalog no. 4106), and Human Neuron Lysate (product code HNL, catalog no. 1526).

Techniques: Sequencing, Western Blot

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: α-Klotho Expression in Human Tissues

doi: 10.1210/jc.2015-1800

Figure Lengend Snippet: α-Klotho protein expression and distribution in human epithelial and reproductive tissues. IHC, positive staining (brown) was found in all the cellular layers of the epidermis (A) and appendage tissue such as hair follicle and sebaceous gland (B). Intestinal expression was primarily found in epithelial cells as illustrated in jejunum (C) and colon (D). In reproductive tissues, positive staining was found in epithelial Sertoli cells (E), testosterone producing Leydig cells (illustrated with white arrows) of the testis (F), and epithelial cells of the prostate gland G). H–K, In mammary tissue (H), endometrium of uterus (I), and endometrium of salpinx (K), the epithelial cell layer was staining strongly for α-Klotho protein; insets are larger magnifications of the epithelial layer. n ≥ 5 for each tissue.

Article Snippet: Cell lysates were obtained from commercially available primary cell cultures (ScienCell Research Laboratories), including: Human Epidermal Keratinocyte Lysate-adult (product code HEKL-a, catalog no. 2116), Human Prostate Epithelial Cell Lysate (product code HPrEpiCL, catalog no. 4416), Human Mammary Epithelial Cell Lysate (product code HMEpiCL, catalog no. 7616), Human Renal Proximal Tubular Epithelial Cell Lysate (product code HRPTEpiCL, catalog no. 4106), and Human Neuron Lysate (product code HNL, catalog no. 1526).

Techniques: Expressing, Staining

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: α-Klotho Expression in Human Tissues

doi: 10.1210/jc.2015-1800

Figure Lengend Snippet: Mass spectrometry characterization of the transmembrane α-Klotho protein in human tissues and cells: extracellular α-Klotho peptide GLFYVDFLSQKD (exon 3). A–D, Representative mass spectrometry spectra (left) and Skyline data (right) confirmed the presence of full-length α-Klotho rh full-length α-Klotho protein (rh-α-Klotho) (A), renal proximal tubular epithelial cells (B), kidney tissue (C), and renal artery (D). E–G, The full-length specific (isoform 1) αKlotho peptide LWITMNEPYTR (exon 4). Representative mass spectrometry spectra (left) and Skyline data (right) confirmed the presence of full-length α-Klotho. E), rh full-length α-Klotho protein (rh-α-Klotho); F, kidney tissue; G, renal artery; and H, neuronal cells.

Article Snippet: Cell lysates were obtained from commercially available primary cell cultures (ScienCell Research Laboratories), including: Human Epidermal Keratinocyte Lysate-adult (product code HEKL-a, catalog no. 2116), Human Prostate Epithelial Cell Lysate (product code HPrEpiCL, catalog no. 4416), Human Mammary Epithelial Cell Lysate (product code HMEpiCL, catalog no. 7616), Human Renal Proximal Tubular Epithelial Cell Lysate (product code HRPTEpiCL, catalog no. 4106), and Human Neuron Lysate (product code HNL, catalog no. 1526).

Techniques: Mass Spectrometry, Targeted Proteomics

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: α-Klotho Expression in Human Tissues

doi: 10.1210/jc.2015-1800

Figure Lengend Snippet: Confirmation of Transmembrane α-Klotho Protein Expression in Human Tissues and Cells

Article Snippet: Cell lysates were obtained from commercially available primary cell cultures (ScienCell Research Laboratories), including: Human Epidermal Keratinocyte Lysate-adult (product code HEKL-a, catalog no. 2116), Human Prostate Epithelial Cell Lysate (product code HPrEpiCL, catalog no. 4416), Human Mammary Epithelial Cell Lysate (product code HMEpiCL, catalog no. 7616), Human Renal Proximal Tubular Epithelial Cell Lysate (product code HRPTEpiCL, catalog no. 4106), and Human Neuron Lysate (product code HNL, catalog no. 1526).

Techniques: Expressing, Recombinant

Journal: Oncotarget

Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

doi: 10.18632/oncotarget.12730

Figure Lengend Snippet: Studies on the prognostic significance of STAT1 and STAT3 in breast cancer

Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

Techniques:

Journal: Oncotarget

Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

doi: 10.18632/oncotarget.12730

Figure Lengend Snippet: No appreciable expression was detected in the negative controls of ph-STAT1 A. and ph-STAT3 B . C-H. show the staining intensity of the STAT1 and STAT3 expression as low (C and D), moderate (E and F), and strong (G and H). Original magnification, 20×. Scale bars = 100 μm (A-F), 10 μm (G and H).

Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

Techniques: Expressing, Staining

Journal: Oncotarget

Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

doi: 10.18632/oncotarget.12730

Figure Lengend Snippet: The relationship between ph-STAT1 and ph-STAT3 tumour cell expression and clinicopathological characteristics (n=384)

Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

Techniques: Expressing

Journal: Oncotarget

Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

doi: 10.18632/oncotarget.12730

Figure Lengend Snippet: A. Total STAT1 tumour cell expression and B. Ph-STAT1 tumour cell expression. C. Total STAT3 tumour cell expression and D. Ph-STAT3 tumour cell expression.

Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

Techniques: Expressing

Journal: Oncotarget

Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

doi: 10.18632/oncotarget.12730

Figure Lengend Snippet: The relationship between clinicopathological characteristics, ph-STAT1 and ph-STAT3 tumour cell expression and cancer specific survival in patients with invasive ductal breast cancer (n=384)

Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

Techniques: Expressing

Journal: Oncotarget

Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

doi: 10.18632/oncotarget.12730

Figure Lengend Snippet: The relationship between clinicopathological characteristics, ph-STAT1 and ph-STAT3 tumour cell expression and cancer specific survival in patients with high grade necrosis (n=201)

Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

Techniques: Expressing

Journal: Oncotarget

Article Title: The relationship between total and phosphorylated STAT1 and STAT3 tumour cell expression, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

doi: 10.18632/oncotarget.12730

Figure Lengend Snippet: Luminal A (n=174, 45%) and luminal B (n=92, 24%) show significant association between high tumour cell expression of ph-STAT3 (n=109, 28%) and improved cancer specific survival. Manipulation is permitted.

Article Snippet: TMA sections were then incubated overnight at 4°C with the primary antibodies as following: total STAT1 (STAT1 (42H3) Rabbit monoclonal antibody, code 9175, Cell Signaling Technology, USA) at a concentration of 1:100; ph-STAT1 (Rabbit PAb to STAT1 phosphoY701, code ab30645, Abcam, Cambridge) at a concentration of 1:150; total STAT3 (STAT3 Rabbit Ab, code 9132L, Cell Signaling Technology, USA) at a concentration of 1:200; Ph-STAT3 (Y705) antibody (P-STAT3 (Y705) Rabbit Ab, code 9131L, Cell Signaling Technology, USA) at a concentration of 1:200.

Techniques: Expressing