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3 protocols using cell culture wares

1

Cell Culture Media Procurement Protocol

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Medium, fetal bovine serum, and penicillin/streptomycin were purchased from Gibco Life Technologies, (Carlsbad, CA). Cell culture wares were purchased from Corning, Inc. (San Mateo, CA). Unless otherwise specified, all other chemicals were purchased from Sigma-Aldrich (St. Louis, MO).
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2

miR-145 Regulation of HER2 and p53

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All chemicals and reagents were purchased from Sigma Adrich Corporation (St. Louis, MO, USA) and cell culture wares were purchased from Corning life sciences (Tewksbury MA, USA). Non-targeting control (catalog number: AM17111) and miR-145 mimics (catalog number: 4464066), Taqman miR-145 probes (Assay id: 002278), High Capacity cDNA Reverse Transcription kit (catalog number: 4368814) and TRIzol reagent (catalog number: AM 9738) were purchased from Life technologies (Carlsbad, CA, USA). The primary antibodies, anti-HER2 (catalog number: 2165S), pAKTSer473 (catalog number: 9271) and anti-p53 (catalog number: 2527) were purchased from Cell Signaling (Danvers, MA, USA) and anti-β–actin (catalog number: A5316) was purchased from Sigma (St. Louis, MO, USA).
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3

Sturgeon Swim Bladder Collagen Extraction

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Type I atelocollagen was extracted from the swim bladder of Bester sturgeon as described previously [28] .
Porcine pepsin (EC 3.4.23.1, 1:10,000, Wako Pure Chemical Industries Ltd., Osaka, Japan) at a concentration of 0.1% was used for removing telopeptides of collagen molecules. For comparison, porcine skin type I collagen (Cellmatrix type I-C, Nitta Gelatin Inc., Osaka, Japan) was used. Cell culture media and penicillin/streptomycin (P/S) solution were obtained from Thermo Fisher Scientific (Waltham, MA).
Hank's balanced salt was obtained from Sigma-Aldrich (St. Louis, MO). Other chemicals were obtained from Wako Pure Chemical Industries Ltd unless otherwise stated. Cell culture wares were from Corning, NY. Cells were obtained from RIKEN Cell Bank (Tsukuba, Japan) and Japanese Collection of Research Bioresources (Tokyo, Japan).
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