Journal: The Journal of Steroid Biochemistry and Molecular Biology

Article Title: Regulation of the calcium-sensing receptor expression by 1,25-dihydroxyvitamin D 3 , interleukin-6, and tumor necrosis factor alpha in colon cancer cells ☆

doi: 10.1016/j.jsbmb.2013.10.015

Figure Lengend Snippet: Schematic illustration of the CaSR promoter region including exon 1A and exon 1B. Position of binding sites for regulatory elements is shown (signal transducer and activator of transcription (STAT), vitamin D response elements (VDRE), nuclear factor kappa B (NF-κB), specificity protein 1 (SP1)), which are critical for 1,25D 3 , TNFα, and IL-6 responsiveness, as well as the CAAT and TATA boxes. Transcription start sites (TSS) 1 and 2 according to were taken as point of reference for positioning the indicated binding sites in the corresponding promoters.

Article Snippet: Cells were incubated either with rabbit polyclonal anti-CaSR antibody (1:100, Anaspec, USA) or mouse monoclonal anti-CaSR antibody (1:200, Abcam, UK) for 1 h at room temperature.

Techniques: Binding Assay

Journal: The Journal of Steroid Biochemistry and Molecular Biology

Article Title: Regulation of the calcium-sensing receptor expression by 1,25-dihydroxyvitamin D 3 , interleukin-6, and tumor necrosis factor alpha in colon cancer cells ☆

doi: 10.1016/j.jsbmb.2013.10.015

Figure Lengend Snippet: Transcriptional regulation of CaSR by 1,25D 3 in colon cancer cell lines. Caco2/AQ and Coga1A cells were treated with 10 nM 1,25D3 for the indicated time points. Bars represent mean ± SEM of 2-3 independent experiments.

Article Snippet: Cells were incubated either with rabbit polyclonal anti-CaSR antibody (1:100, Anaspec, USA) or mouse monoclonal anti-CaSR antibody (1:200, Abcam, UK) for 1 h at room temperature.

Techniques:

Journal: The Journal of Steroid Biochemistry and Molecular Biology

Article Title: Regulation of the calcium-sensing receptor expression by 1,25-dihydroxyvitamin D 3 , interleukin-6, and tumor necrosis factor alpha in colon cancer cells ☆

doi: 10.1016/j.jsbmb.2013.10.015

Figure Lengend Snippet: Effect of 1,25D 3 , TNFα, and IL-6 on CaSR expression. (A and B) mRNA expression of Caco2/AQ and Coga1A cells assessed by real time qRT-PCR. Data were log transformed to achieve normal distribution, then subjected to one way ANOVA and corrected with Tukey's posttest for multiple comparisons. Bars represent mean ± SEM of 2-3 independent experiments, asterisks above bars indicate statistically significant changes compared with control. * p < 0.05, ** p < 0.01. (C and D) Immunofluorescence staining of the CaSR protein (red) and nuclear staining (blue). Scale bar was 50 μm.

Article Snippet: Cells were incubated either with rabbit polyclonal anti-CaSR antibody (1:100, Anaspec, USA) or mouse monoclonal anti-CaSR antibody (1:200, Abcam, UK) for 1 h at room temperature.

Techniques: Expressing, Quantitative RT-PCR, Transformation Assay, Control, Immunofluorescence, Staining

Journal: Journal of Bone and Mineral Research

Article Title: Mapping of Human Autoantibody Binding Sites on the Calcium-Sensing Receptor

doi: 10.1359/jbmr.090703

Figure Lengend Snippet: Synthetic peptide ELISAs with patient and control sera. Patient and control sera were analyzed for antibody binding to synthetic peptides in an ELISA format, as detailed in Materials and Methods. The Ab indices are shown for APS1 ( n = 14) and AHH ( n = 3) patient sera and for control ( C ) ( n = 10) serum samples analyzed in ELISA experiments against synthetic CaSR peptides 41–69, 114–126, 214–238, 344–358, and 374–391.

Article Snippet: Anti-CaSR rabbit polyclonal antibody against a synthetic peptide corresponding to amino acids 12–27 of the rat CaSR was purchased from Alexis Biochemicals (Nottingham, UK).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay