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Sterilized water

Manufactured by Merck Group

Sterilized water is a laboratory-grade purified water product. It is processed to remove impurities and microorganisms, ensuring a high level of purity and sterility for use in various laboratory applications.

Automatically generated - may contain errors

2 protocols using sterilized water

1

Micropatterned Hydrogel Fabrication via Soft Lithography

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Polylactic acid (PLA, 3 mm granule, Mw 193.3 kg/mol) was purchased from GoodFellow;
chloroform and dexamethasone (Pharmaceutical Secondary Standard, Supleco)
were purchased from Sigma-Aldrich. A poly(dimethylsiloxane) (PDMS)
kit from Sylgard was used to create the PDMS stamps. The PDMS kit
(Sylgard 184, Dow-Corning, USA) was used for PDMS stamp fabrication
by the standard soft lithography technique using a silica mold.29 (link) The mold contained an array of 11 × 11
× 22 μm rectangular pillars on an area 20 × 20 mm2. It was fabricated by cryogenic reactive ion etching of a
silica wafer with a chromium mask made by lift-off lithography. 2-Hydroxyethyl
methacrylate (HEMA) (97%, Sigma-Aldrich), ethylene glycol dimethacrylate
(EGDMA) (98%, Sigma-Aldrich), and photoinitiator (Irgacure 2959; Ciba)
were used for the hydrogel solution. Phosphate buffer saline (PBS)
(tablet, Sigma-Aldrich) and sterilized water (Sigma-Aldrich) were
used to make the PBS solution. To label the samples for confocal microscopy,
Nile red (Sigma-Aldrich), 5(6)-carboxyfluorescein (CF) (Sigma-Aldrich),
and dexamethasone fluorescein (ThermoFisher Scientific) were used.
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2

Luciferase Reporter Assay for CircRNA-miRNA Interactions

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Wild‐type and mutant reporter plasmids including circ_0087199 WT, TLR4 3′UTR WT, circ_0087199 WT, and TLR4 3′UTR WT were built using pmirGLO vector (Miaoling biotechnology) following the prediction of binding sites of miR‐527 for circ_0087199 and TLR4 through circRNAs interactome and Targetscan online databases. These plasmids were transfected into PDLCs with miR‐527 mimics or mimic control, and the culture medium was removed from the cells into which the reporter plasmids had been transferred. Lysis buffer diluted with sterilized water (Sigma) was used to lysate cells. After centrifugation, the supernatant was utilized for luciferase activity analysis as instructed in the manual of a commercial dual luciferase reporter assay kit (Life‐iLab biotech).
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