Journal: Endocrinology
Article Title: Thymic PTH Increases After Thyroparathyroidectomy in C57BL/KaLwRij Mice
doi: 10.1210/en.2017-03083
Figure Lengend Snippet: Thymus-derived PTH increases after TPTX. After serum PTH returned to a pre-TPTX level, TPTX and control mice were euthanized, and parathyroid and thyroid (Thyroid+Parathyroid), liver, and thymus tissues were removed. Harvested tissues were washed extensively in PBS and homogenized in T-PER buffer, and proteins were immunoprecipitated with goat anti-mouse PTH antibodies, including anti-mouse PTH(1-12) and anti-mouse PTH(53-84) or goat IgG as an isotype control. (A) PTH was detected in whole cell lysates (WCL) of the parathyroid+thyroid, after immunoprecipitation from the parathyroid+thyroid, and in the thymus (indicated by arrows). PTH protein was not detected in the liver. (B) Thymi from TPTX mice contained more PTH than thymi from nonoperated (control) mice. (C) Immunohistochemistry showing increased PTH protein production in the thymus after TPTX. Thymus sections were prepared and incubated with anti–PTH(1-12) antibody. The antibody was visualized with 3,3′-diaminobenzidine and counterstained with methyl green. The representative PTH signals are indicated by yellow arrows. (D) PTH-positive cells were enumerated in at least four sections from each group and plotted as mean ± standard error. Student t test was performed. ***P < 0.001. IP, immunoprecipitated; Iso, isotype control antibody; PT, parathyroid glands; PTH*, parathyroid hormone antibody.
Article Snippet: Tissue lysates were incubated with 1 μg goat anti-mouse PTH antibodies, including anti-mouse PTH(1-12) [catalog no. 20-2320; Research Resource Identifier: AB_2721076 ; Quidel, San Diego, CA] and anti-mouse PTH(53-84) (catalog no. 20-2310; Research Resource Identifier: AB_2721077 ; Quidel) or with 1 μg goat immunoglobulin G (IgG; catalog no. ab37373; Abcam, Cambridge, MA) as an isotype control for 30 minutes at 4°C on a rotary shaker.
Techniques: Derivative Assay, Immunoprecipitation, Immunohistochemistry, Incubation
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