Journal: International Journal of Clinical and Experimental Pathology

Article Title: Relationship between CaSRs and LPS-injured cardiomyocytes

doi:

Figure Lengend Snippet: Expression of CaSR in cardiomyocytes as indicated by western blot (n=8). A basal level of CaSR was detected in the control group. This served as a baseline for the comparison of fold increases in other groups. The expression of CaSR in the LPS group was higher than that in the control group. Pretreatment with SC-211006 further increased CaSR expression. However, SC-207394 was found to inhibit the expression of CaSR. **P<0.05 vs. the control group, ▲▲P<0.05 vs. the LPS group.

Article Snippet: Reagents LPS was obtained from Sigma (St. Louis, USA), SC-211006 and SC-207394 were obtained from Santa Cruz Biotechnology, anti-CaSR antibody (product number: ACR-004) was purchased from Alomone Labs (Jerusalem, Israel), anti-GRP78 antibody (product number: 3158-1) was purchased from Epitomics (Burlingame, USA), and anti-LC3B antibody (product number: TDY357), anti-CytC antibody (product number: TDY378) and anti-Bcl-2 antibody (product number: TDY061) were obtained from Tian De Yue (Beijing, China).

Techniques: Expressing, Western Blot, Control, Comparison

Journal: bioRxiv

Article Title: Differential activation of parathyroid and renal Ca 2+ -sensing receptors underlies the renal phenotype in autosomal dominant hypocalcemia 1

doi: 10.1101/2021.02.01.429170

Figure Lengend Snippet: Ca 2+ homeostasis in a patient with ADH1 and relationship between CaSR abundance and CaSR activity. Treatment of a patient with ADH1 in order to normalize blood Ca 2+ levels were associated with the occurrence of hypercalciuria when the patient was normocalcemic ( n =6-19 individual measurements per cluster) (A). In addition, fractional urinary excretion of Ca 2+ (FE Ca 2+ ) was increased after treatment leading to normocalcemia ( n =6-19 individual measurements per cluster) (B). Plasma intact PTH levels (iPTH) were suppressed during all collection periods ( n =6-19 individual measurements per cluster) (C). Medullary calcinosis, a complication of treatment-induced hypercalciuria in ADH1, was observed by ultrasound (D). Immunohistochemistry revealed a marked immunoreactivity of CaSR in the parathyroid (E). CaSR immunoreactivity was also detected in the thick ascending limb of Henle’s loop in the kidney, albeit at a lower intensity (F). After transfection of HEK293 cells with different amounts of a construct encoding the human CaSR (hCaSR) and a fixed amount of a CLDN14-promoter reporter construct, higher reporter signal was obtained after transfection of cells with a higher amount of construct ( n =4 independent experiments) (G). The increased hCaSR expression was confirmed by western blotting with an anti-CaSR antibody (H) and with an anti-C-myc antibody (I). Representative immunohistochemical images and western blots are shown; arrows indicate dimeric CaSR signal. Data are shown as mean ± SEM. * P < 0.05 by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: For the immunohistochemical analyses of the parathyroid and renal cortex, sections were processed as described above and stained using a rabbit polyclonal antibody against the CaSR (PB9924, Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Activity Assay, Clinical Proteomics, Immunohistochemistry, Transfection, Construct, Expressing, Western Blot, Immunohistochemical staining

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