Dehydrated alcohol

Hyaluronic acid (Mw ≈ ~800 kDa, F1177), collagen (11179179001), plasminogen (341578), Dulbecco’s modified Eagle’s medium (DMEM, D0819), 4′6-diamidino-2′-phenylindole dihydrochloride (DAPI, 10236276001), phosphate buffered saline (PBS, pH 7.4, P4417), bovine serum albumin (BSA, A3858), paraffin (327204), and dehydrated alcohol (1012772) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS, 16140071), penicillin/streptomycin (P/S, 15070063), trypsin (0.25%, 15050065), primary antibody to CD31 (37-0700), Rhodamine Phalloidin (R415) were obtained from Gibco (Grand Island, NY, USA). Anti-alpha (ɑ)-smooth muscle actin (ɑ-SMA, ab5694) and Cell Counting Kit-8 (CCK-8, ab228554) were purchased from Abcam (Cambridge, UK). The 4% paraformaldehyde (C104190), Masson’s trichrome (MTC), Verhoeff-Van Gieson (VVG, GP1035), Triton X-100 (WGT8200), hematoxylin eosin (H&E, G1004, G1002), secondary antibodies, Alexa Fluor-488, or Cy3-conjugated anti-mouse or anti-rabbit immunoglobulin-G (IgG) for fluorescence staining were obtained from Servicebio Science and Technology Co., Ltd (Wuhan, China). Hexafluoroisopropanol (HFIP, 920-66-1) was purchased from Aladdin (Shanghai, China). Mouse PAECs (SNP-M016) and SMCs (SNP-M015) were obtained from Sunncell Bioscience Inc. (Wuhan, China).

A cell invasion assay was performed using 24-well Transwell chambers (Corning, Inc.) and the inserts were coated with 50 μl Matrigel^® (Dilution, 1:8 with DMEM; BD Biosciences, Franklin Lakes, NJ, USA). Normal U251 and RR-U251 cells treated with CFL1-siRNA and pcDNA3.1-CFL1 were cultured in six-well plates. Following radiotherapy, the monolayer cells were trypsinized and transferred to the upper Matrigel chamber in 100 μl serum-free DMEM at a density of 1×10⁵/ml. DMEM supplemented with 15% fetal bovine serum (Gibco, Invitrogen Life Technologies) was added to the lower chamber as the chemoattractant. Following incubation for 24 h, cells remaining in the upper chamber were removed using cotton swabs, while invaded cells were fixed using dehydrated alcohol (Sigma-Aldrich), stained with crystal violet (Sigma-Aldrich) and then counted under a microscope (Axiovert 40 CFL). Images were captured in five randomly selected fields for each well (magnification, ×100). Three separate experiments were performed.

Specifically, 8-OHdG, bovine serum albumin (BSA), citric acid, diethylenetriamine (EDTA), (3-glycidyloxypropyl)trimethoxysilane (GPMS), glutaraldehyde, methylbenzene, chloroauric acid, 3-mercaptopropionic acid (MPA), zinc nitrate hexahydrate (Zn(NO₃)₂·6H₂O), methanol (99.8%), sodium chloride, potassium chloride, calcium chloride, acetone, N,N-dimethylformamide (DMF), sodium borohydride (NaBH₄, 99.99%), and dehydrated alcohol were obtained from Sigma Aldrich (St. Louis, Missouri (Mo), USA). NaCl, KCl, CaCl₂, MgCl₂, thymine, cytosine, adenine, guanine, and hydrogen peroxide were ordered from ALADDIN Reagent (Shanghai, China). Nanoporous alumina membranes were purchased from Whatman (Boston, Massachusetts (Ma), USA). Alongside, 8-OHdG antibody was bought from Abcam (Cambridge, UK). In addition, 2-methylimidazole (2-MeIM, 99%), 1-dodecanethiol (DDT, ≥98%), hexadecyltrimethyl ammonium bromide (CTAB, 98%), silver nitrate (AgNO₃, ≥99.0%), 11-mercaptoundecanoic acid (MUA, 95%), and 4-nitrophenol (4-NP, ≥99.5%) were purchased from Sigma Aldrich (St. Louis, Missouri (Mo), USA). The chemicals were used as received without further purification.