Sc 20736

Rabbit anti-Ki67 (1:300; ab15580, Abcam), rabbit anti-Mucin 2 (1:300; ab272692, Abcam), rabbit anti-lysozyme (1:300; ab108508, Abcam), rabbit anti-chromogranin A (1:300; ab15160, Abcam), mouse anti–E-cadherin (1:1000; 610182, BD Biosciences), mouse anti–α-catenin (1:500; 610194, Biosciences), mouse anti–β-catenin (1:500; 610154, Biosciences), rabbit anti-BMPR1A (1:1000; sc-20736, Santa Cruz Biotechnology), rabbit anti-Smad4 (1:1000; sc7154, Santa Cruz Biotechnology), rabbit anti-Olfm4 [1:300; 39141, Cell Signaling Technology (CST)], mouse anti-vimentin (1:500; 550513, BD Biosciences), rabbit anti–IL-1α (1:50; ab300499, Abcam), rabbit anti–IL-1α (1:50; 16765-1-AP, Proteintech), mouse anti–IL-1β (1:200; sc-52012, Santa Cruz Biotechnology), rabbit anti-MOT10 (1:1000; YT7177, ImmunoWay), rabbit anti-CHP2 (1:1000; YT0915, ImmunoWay), rabbit anti–p-Smad1/5 (1:100; 9516, CST), mouse anti-Ki67 (1:200; 9449, CST), and rabbit anti-Gli1 (1:80; 3538, CST).

Paraffin-embedded P1 lower jaws (n = 5) were sliced to a thickness of 8 μm. Selected sections were then soaked in L.A.B. Solution (Polysciences, PA, USA) for 5 min for antigen activation, followed by incubation in 5% BSA/PBS for 1 h. After incubation with the primary antibodies for BMPRIA (sc-20736; Santa Cruz Biotechnology, CA, USA), BMPRIB (sc-25455; Santa Cruz Biotechnology), or BMPRII (MABD171; Merck Millipore, MA, USA), expression was detected using Alexa488- or Alexa594-conjugated secondary antibodies (Life Technologies). A fluorescence microscope (BZ-8000, KEYENCE Co, Osaka, Japan) was used for image analysis. Images were prepared using a BZ analyzer (KEYENCE) and Photoshop (Adobe Systems, CA, USA).

BMPR1A (1:20, sc20736; Santa Cruz); BRACHYURY/T (1:400, AF2085; R&D); Clover (1:600, EMU101; Kerafast); OCT4 (1:800, sc8628; Santa Cruz); pSMAD1/5 (1:800, 13820s; Cell Signaling); TGFBR1 (1:20, sc9048; Santa Cruz); ZO-1 (1:100, 33-9100; Thermo Fisher); ZO-1-FITC (1:100, 33-9111; Thermo Fisher).