P38 mapk

The following items were purchased from the indicated sources: BCA protein assay kit was purchased from Dalian Meilun Biotechnology (Dalian, China). primary antibodies against E-cadherin (rabbit, #40860, Signalway Antibody, MD, USA), N-cadherin (rabbit, A5598, Bimake, Houston, TX, USA), Vimentin (rabbit, A5862, Bimake, Houston, TX, USA), phospho-ERK1+2 (rabbit, bs-1522R, Bioss, China), phospho-JNK1+2+3 (rabbit, bs-1640R, Bioss, Beijing, China), phospho-p38 MAPK (rabbit, bs-0636R, Bioss, Beijing, China), ERK1+ERK2 (rabbit, bs-2637R, Bioss, Beijing, China), JNK1+JNK2+JNK3 (rabbit, bs-2592R, Bioss, Beijing, China), p38 MAPK (rabbit, bs-0637R, Bioss, Beijing, China), GAPDH (rabbit, AP0066, Bioworld, MN, USA), Goat Anti-Rabbit IgG(H+L)/HRP (rabbit, bs-40295G-HRP, Bioss, Beijing, China) and Goat Anti Rabbit IgG (H&L)-Alexa Fluor 488 (rabbit, ImmunoWay, Plano, TX, USA).

All samples including the maxilla and lymph node were firstly fixed in 4% paraformaldehyde for 24 h. Maxilla were then decalcified in 10% ethylene diamine tetraacetic acid for 3 months before being embedded in paraffin. Serial Sect. (3 μm) of maxilla samples were cut from the mesial of the second molar to the distal. Deparaffinized sections were stained with hematoxylin and eosin and immunohistochemistry staining (MXB Biotechnologies) according to the manufacturer’s protocol. Primary antibodies used were GATA-3 (1:50, proteintech), IFN-γ (1:2000, Bioss), IL-4 (1:200, proteintech), OPG (1:500, proteintech), ALP (1:2000, proteintech), RANK (1:2000, Bioss), ERK1 (1:500, Bioss), and p38 MAPK (1:50, Bioss). Negative controls were used to test immunostaining specificity by replacing the primary antibody with phosphate buffer saline.

Antibodies from Affinity Biosciences: p-Smad1/5/8 (1:1000, #AF8313), Smad1/5/8 (1:1000, #AF0614), PPARγ (1:1000, #bs-4590R), UCP1 (1:1000, #72298). Antibodies from Beyotime: HA tag (1:1000, #AH158), Flag tag (1:1000, #AF519). Antibodies from Sangon Biotech: p-ACC (1:1000, #D155180), ACC (1:1000, #D155300). Antibodies from Abcam: BMP8A (1:1000, #154373). Antibodies from ZenBio: p-AMPK (1:1000, #R26252), AMPK (1:1000, #380431), PGC-1α (1:1000, #381615). Antibodies from CWBIO: goat anti-Rabbit IgG HRP secondary antibody (1:4000, #CW0103S), goat anti-Mouse IgG HRP Conjugated (1:4000, # CW0102). Antibodies from Bioss: p38 MAPK (1:1000, #bs-0637R), p-p38 MAPK (1:1000, #bs-2210R), β-actin (1:2000, #bs-0061R), p-JNK (1:1000, #bs-1640R), JNK (1:1000, #bs-2592R), C/EBPα (1:1000, #AF6333), p-Smad2/3 (1:1000, #AF3367), Smad2/3 (1:1000, #AF6367), p-ERK1/2 (1:1000, #AF1015), ERK1/2 (1:1000, #AF0155), p-p65 (1:1000, #AF2006), p65 (1:1000, #AF5006), p-IKKα/β (1:1000, #AF3013), IKKα/β (1:1000, # AF6014).