Hmgb1 antibody 10829 1 ap

HMGB1 antibody (10829-1-AP) was purchased from Proteintech (Chicago, IL, USA). PDGFR-α (3164S), PDGFR-β (3169S) and phospho-specific (2992S, 3166S) antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). β-Actin (sc-47778) and Lamin B1 (sc-374015) antibodies were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). NE-PERTM reagents (78835) were purchased from Thermo Fisher Scientific (Rockford, IL, USA). Puredown protein G-agarose (P9202-050) was purchased from GenDEPOT (Barker, TX, USA). IP lysis buffer (87787) was purchased from Thermo Fisher Scientific. Various growth factor receptor inhibitors such as AG1024 (65678-07-1; an IGF receptor inhibitor), AG1295 (71897-07-9; a PDGF receptor inhibitor), AG1478 (175178-82-2; an EGF receptor inhibitor), and PD173074 (219580-11-7; an FGF receptor inhibitor) were purchased from Calbiochem (La Jolla, CA, USA). Recombinant PDGF-AA (221-AA) and-DD (1159-SB) were purchased from R&D Systems Inc. (Minneapolis, MN, USA). Horseradish peroxidase (HRP)-conjugated IgG secondary antibodies were purchased from Santa Cruz Biotechnology Inc.

The human airway biopsy and mouse left lungs were paraffin-embedded, and 5-μm sections were cut. Sections were used for hematoxylin and eosin (H&E) staining, periodic acid-Schiff (PAS) staining and immunohistochemical examination. For immunofluorescence staining, bronchial biopsy sections underwent antigen retrieval and were incubated with mouse monoclonal RUNX2 antibody (sc-390715 X, 1:1000 dilution, Santa Cruz Biotechnology, USA) and rabbit polyclonal HMGB1 antibody (10829-1-AP, 1:200 dilution, proteintech, China), then incubated with Cy3 goat anti-mouse secondary antibody (ServiceBio, China) and FITC goat anti-rabbit secondary antibody (ServiceBio, China). Nuclei were stained with DAPI. Photographs were taken by using a fluorescence microscope (NIKON Eclipse ci, Japan). For PAS staining, lung sections were stained with PAS (Goodbio technology, China) for detection of mucus. The number of PAS-staining-positive cells was counted in 10 random fields for each lung section at ×200 magnification. Observers who were blinded to the clinical status of the subjects counted numbers of eosinophils/mm² submucosa as previously described [28 (link)].

Zn (NO3)2•6H2O and 2-MeIM were purchased from Aladdin Holdings Group Co., Ltd (Shanghai); PEG-FA 2000, ICG, and FITC from Rixi Biotechnology Co., Ltd. (Xi’an); BMS202 (HY-19745), STF62247 (HY-100746), 3-MA (HY-19312), and Chloroquine (HY-17589A) from MedChemExpress (Shanghai); ethanol from Guangzhou Chemical Reagent Factory.
LC3B antibody (L7543) was purchased from Sigma; ATG5 antibody (T55766) from Abmart; p-mTOR antibody (Ser172) (#2971S) from CST; HMGB1 antibody (10,829-1-AP) from Proteintech; HRP-Goat anti-rabbit IgG(H + L) (E030120-01) from Earthox; CD4 antibody (bs-0647R), CD8 antibody (bs-4791R) from Bioss;p62 antibody (ab109012), mTOR antibody (ab137133), Calreticulin antibody (ab92516), Ki67 antibody (ab16667), Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077), Goat anti-rabbit IgG H&L (Alexa Fluor® 555) (ab150078), and DAPI Staining Solution (ab228549) from Abcam.