Mandibles of 6- to 8-week-old WT and Orai1K14 and Orai2−/− mice were extracted and cleaned of soft tissues, dehydrated at 4°C in ethanol series, and embedded in polymethyl methacrylate (PMMA) resin, and the erupted incisor was cut with a circular diamond saw about 2 to 3 mm from the tip and was polished. Detailed protocols were previously reported (6). BSE imaging was performed to assess mineralization differences in a Zeiss EVO-50 microscope without a conductive coating in the following settings: 50 Pa, 15 kV, and 200 pA. WT and corresponding Orai1K14 and Orai2−/− samples were analyzed in parallel, setting contrast and brightness arbitrarily to the range of WT samples. To determine microstructural differences, samples were acid-etched (phosphoric acid, 37% for 5 s), rinsed in deionized water, and imaged in BSE. FE-BSE imaging was conducted in a Zeiss Gemini 300 in nano-VP mode using BSD1 detectors at a 7.4-mm working distance. A total of three Orai1K14 and three Orai2−/− mice were analyzed plus a corresponding three WT mice per group.

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