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3 protocols using hmgb2

1

Adipocyte Differentiation Protocol

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Mouse embryo fibroblast 3T3-L1 (ATCC®CL173) and normal human primary subcutaneous preadipocytes (ATCC® PCS210010) cells were purchased from American Type Culture Collection (ATCC, Manassas, VA). Dulbecco’s modified Eagle’s medium high glucose (DMEM), penicillin/streptomycin and L-glutamine were purchased from Mediatech, Inc. (Manassas, VA). PPARγ (Cat# 2443S), C/EBPα (Cat# 2295S), FAS (Cat# 4233S), HMGB2 (Cat# 14163S), AMPKα (Cat# 5832S) and phospho(p)-AMPKα (Cat# 50081S-Thr172), Akt1(Cat# 75692S) and p-Akt1 (Cat# 9018S-Ser473) antibodies were from Cell Signaling Technology (Boston, MA). GAPDH (Cat# G9545) antibodies were from Sigma-Aldrich (St. Louis, MO). All secondary antibodies (Cat# 305-035-045) were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA). All other chemicals were obtained from Sigma-Aldrich unless otherwise stated.
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2

Immunoblotting Protocol for Protein Analysis

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Cells were lysed, and immunoblotting was performed as previously described (Bianco and Mohr, 2017 (link)). The antibodies used in this study were as follows: TIF-IA (Bethyl Laboratories; A303-065A) UBF (Santa Cruz; SC-13125), Akt (Cell Signaling; 9272), Fibrillarin (Santa Cruz; sc-166001), UL44 (Virusys; CA006), pp28 (Virusys; CA004-100), IE1/IE2 (Millipore; MAB810), HMGB2 (Cell Signaling Technology; 14163), USP18 (Cell Signaling Technology; 4813), IFIT2 (Proteintech; 12604–1-AP), TFIIIA (Bethyl; A303-621A), ISG15 (Proteintech; 15981–1-AP), RIG-I (Proteintech; 20566), MDA5 (Proteintech; 21775–1-AP), UBA7 (Cell Signaling Technology; 69023), actin (Cell Signaling Technology; 3700), BrdU (Sigma; B2531), p53 (Cell Signaling Technology; 9282).
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3

Adipocyte Differentiation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mouse embryo fibroblast 3T3-L1 (ATCC®CL173) and normal human primary subcutaneous preadipocytes (ATCC® PCS210010) cells were purchased from American Type Culture Collection (ATCC, Manassas, VA). Dulbecco’s modified Eagle’s medium high glucose (DMEM), penicillin/streptomycin and L-glutamine were purchased from Mediatech, Inc. (Manassas, VA). PPARγ (Cat# 2443S), C/EBPα (Cat# 2295S), FAS (Cat# 4233S), HMGB2 (Cat# 14163S), AMPKα (Cat# 5832S) and phospho(p)-AMPKα (Cat# 50081S-Thr172), Akt1(Cat# 75692S) and p-Akt1 (Cat# 9018S-Ser473) antibodies were from Cell Signaling Technology (Boston, MA). GAPDH (Cat# G9545) antibodies were from Sigma-Aldrich (St. Louis, MO). All secondary antibodies (Cat# 305-035-045) were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA). All other chemicals were obtained from Sigma-Aldrich unless otherwise stated.
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