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. 2015 Aug 6;97(2):302–310. doi: 10.1016/j.ajhg.2015.05.021

Figure 3.

Figure 3

THOC2 Immunofluorescence Staining Is Significantly Reduced in THOC2 p.Leu438Pro Fibroblasts

Fibroblasts seeded on coverslips precoated with BD BioCoat were fixed with 4% paraformaldehyde for 10 min at room temperature and permeabilized with 0.2% Triton X-100 for 10 min at room temperature. Cells were then incubated with rabbit anti-THOC2 antibody (303-630A; Bethyl Laboratories) at 500 ng/ml in 10% horse serum in 1 × PBS overnight at 4°C and then in a 1:2,500 dilution of Alexa Fluor 488 donkey anti-rabbit IgG (H+L) antibody in 10% horse serum in 1 × PBS for 1 hr at room temperature and mounted in ProLong Gold Antifade Mountant with DAPI. Cells were imaged with an Axiocam camera (AxioCam MRm). Scale bars represent 20 μm. See also Figure S4.