Journal: Stem cells (Dayton, Ohio)

Article Title: Intranuclear Actin Structure Modulates Mesenchymal Stem Cell Differentiation

doi: 10.1002/stem.2617

Figure Lengend Snippet: Intranuclear actin is required for both Cyto D-induced adipogenesis and osteogenesis. (A, B): Reverse transcriptase polymerase chain reaction (RT-PCR) from control and Cyto D-treated mesenchymal stromal cells (MSCs) demonstrate expression of adipogenic (Fabp4, Adipoq, and Pparg) and osteogenic (Alpl, Sp7, and Bglap) genes in growth medium; *, p < .05 (A) or in adipogenic medium (B). (C): Western blot analysis for MSCs in adipogenic medium. (D): Importin-9 silencing analyzed by RT-PCR for ±Cyto D-treated cells. (E): Western blot and (F) RT-PCR analysis for ±Cyto D-treated cells ±importin-9 silencing. Abbreviations: A, adipogenic medium; CTL, control; Cyto D, cytochalasin D; O, osteogenic medium; siCTL, control siRNA; silpo9, siRNA targeting importin 9.

Article Snippet: After blocking, primary antibody was applied overnight at 4°C including antibodies against Bglap, Pparg, PARP1 (Cell Signaling, Danvers Mass); Runx2, mDia2, MKL-1, importin-9, Arp3 (Abcam), mDia1 (BD), LDHA (Millipore, St Louis, Mo), Fabp4 (ProSci, Poway, CA), actin, beta-tubulin (Santa Cruz, Dallas TX), Adipoq (Affinity Bioreagents, Golden, CO).

Techniques: Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot

Journal: Diabetes

Article Title: Identification of a Loss-of-Function Mutation in Ube2l6 Associated With Obesity Resistance

doi: 10.2337/db12-1054

Figure Lengend Snippet: Decreased adipocyte differentiation in BALB/c SV preadipocytes. Neutral lipid staining with oil red O in adipocytes on day 7 postdifferentiation induction in ( A ) BALB/c and B6 cells or in ( B ) BALB.2 CC and BALB.2 BC cells. Determination of the number of oil red O–stained adipocytes per field in ( C ) BALB/c and B6 cells or in ( D ) BALB.2 CC and BALB.2 BC cells ( n = 5). mRNA expression analysis by quantitative RT-PCR of Pparγ , Ap2 , Atgl , and Hsl normalized to Actin in ( E ) BALB/c and B6 cells or in ( F ) BALB.2 CC and BALB.2 BC cells ( n = 5). Unpaired t tests were performed. ** P < 0.005. *** P < 0.0005.

Article Snippet: Immunoblots were incubated with primary antibodies against hormone-sensitive lipase (HSL), peroxisome proliferator–activated receptor PPARγ, ATGL, β-actin (Cell Signaling), AP2 (R&D), and UBE2L6 (Santa Cruz).

Techniques: Staining, Expressing, Quantitative RT-PCR

Journal: The FASEB Journal

Article Title: Sphingosine 1-phosphate is a ligand for peroxisome proliferator-activated receptor-γ that regulates neoangiogenesis

doi: 10.1096/fj.14-261289

Figure Lengend Snippet: Treatment of ECs and U937 cells with PPARγ results in differential gene expression. Expression of (A) PGC1β, (B) PAI-1, (C) CD36, and (D) aP2 in ECs and U937 cells in response to 16-h treatment with 5 μM S1P, 5 μM LPA, and 5 μM rosiglitazone (TZD) was determined by qPCR. Gene expression was normalized to CYCA. Data are expressed as mean relative fold changes + sem with vehicle (depicted as dotted line) (n = 6). *P < 0.05 and **P < 0.001 determined by unpaired t test compared to vehicle. E) Protein expression in ECs in response to 5 µM S1P for 16 h was determined using SDS-PAGE and immunoblotting. Representative blot from n = 3.

Article Snippet: Protein expression was detected using the following antibodies: anti-PGC1β (H-300); anti-PPARγ (H-100; Santa Cruz Biotechnology); anti-tubulin (Abcam, Cambridge, United Kingdom); anti-His (Abcam); anti-Flag M2 and anti-PAI-1 (Santa Cruz Biotechnology); anti-CD36 (Abcam); anti-aP2 (A-FABP; Santa Cruz Biotechnology); and anti-β actin (EMD Millipore, Billerica, MA, USA).

Techniques: Expressing, SDS Page, Western Blot