Journal: Kidney international

Article Title: Adipose tissue explants and MDCK cells reciprocally regulate their morphogenesis in coculture.

doi: 10.1038/ki.2010.68

Figure Lengend Snippet: Figure 8 | Adipokine treatment. Effects of leptin (b–e), its antibody (insets in b–e), and combination of leptin and adiponection (f) on morphology of Madin–Darby canine kidney (MDCK) cells at 7 days in culture. (a) (Control), MDCK cells without leptin and adiponectin treatment show a flat shape with cytoplasm and flattened nuclei. (b and c) 20 (b) or 50 ng/ml (c) leptin does not affect the morphology of MDCK cells. (d and e) 100 (d) or 1000 ng/ml (e) leptin allows MDCK cells to develop a tall and large columnar shape, with clear cytoplasm and vertically and basally situated nuclei. The leptin-induced morphology is similar to the adipose tissue fragment (ATF)- induced morphology of MDCK cells. Leptin antibody (L-ab, 25 mg/ml) abolishes the MDCK cell morphology induced by 100 (inset in d) or 1000 ng/ml leptin (inset in e), although the antibody does not affect the morphology of cells at the concentration of 20 (inset in b) or 50 ng/ml (inset in c) leptin. (f) Adiponectin (10 mg/ml) abolishes the MDCK cell morphology induced by 1000 ng/ml leptin.

Article Snippet: To examine the specificity of a leptin-induced effect on the morphology of MDCK cells, 25 mg/ml of rabbit polyclonal leptin antibody against rat leptin (BioVendor LLC, Candler, NC, USA) was added to the 20, 50, 100, or 1000 ng/ml leptin-treated cultures of MDCK cells alone.

Techniques: Control, Concentration Assay

Journal: Kidney international

Article Title: Adipose tissue explants and MDCK cells reciprocally regulate their morphogenesis in coculture.

doi: 10.1038/ki.2010.68

Figure Lengend Snippet: Figure 9 | Adiponectin and leptin production of adipose tissue fragments (ATFs) in the absence or presence of Madin–Darby canine kidney (MDCK) cells in the supernatants of outer and inner dishes at 7 days in culture. In the cultures of ATFs alone, adiponectin and leptin are detected. Their concentration is not significantly different between the outer and inner dishes (P40.05). In the supernatants of outer dishes, adiponectin concentrations in the cocultures of ATFs and MDCK cells are significantly higher than those in the cultures of ATFs alone (Po0.05), whereas leptin concentrations are not significantly different between cultures of ATFs with and without MDCK cells (P40.05). In cocultures of ATFs and MDCK cells, adiponectin (Po0.01) and leptin (Po0.05) concentrations in the inner dishes were significantly lower than those in the outer dishes.

Article Snippet: To examine the specificity of a leptin-induced effect on the morphology of MDCK cells, 25 mg/ml of rabbit polyclonal leptin antibody against rat leptin (BioVendor LLC, Candler, NC, USA) was added to the 20, 50, 100, or 1000 ng/ml leptin-treated cultures of MDCK cells alone.

Techniques: Concentration Assay

Journal: Scientific Reports

Article Title: Mitochondrial sub-cellular localization of cAMP-specific phosphodiesterase 8A in ovarian follicular cells

doi: 10.1038/s41598-019-48886-8

Figure Lengend Snippet: Immunolocalization in mitochondrial isolated fractions from granulosa cells by immunofluorescence using anti-P450scc (green, B , United States Biological), anti-PDE8A (green, E , Proteintech) and MitoTracker (orange, A , D ). Merge signals are shown in ( C , F ). The signal was calibrated with non-specific IgG. Magnification is shown as a bar of 10 µm.

Article Snippet: The samples were washed twice in PBS solution, centrifuged at 14,000 × g for 5 min to eliminate residual markers, fixed with 4% (w/v) PFA in 0.2% Triton X-100 for 10 min in the dark at room temperature, blocked with 5% BSA in PBS for 1 h, then kept at 4 °C overnight with PDE8A polyclonal antibody (Proteintech product number 13956-1-AP, 1:500 in PBS) or porcine P450scc polyclonal antibody (United States Biological, Salem, MA, product number 362774, 1:100 in PBS).

Techniques: Isolation, Immunofluorescence

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