Results: 5

1.
Figure 4

Figure 4. BME suppresses expression of inflammatory cytokines.. From: Beta-Mecaptoethanol Suppresses Inflammation and Induces Adipogenic Differentiation in 3T3-F442A Murine Preadipocytes.

Messenger RNA expression of selected cytokines on day 1, 4, and 10 after incubation with BME at graded concentration of BME. Results are means +/− SE (N = 3–6). Bars denoted with non-identical alphabets are statistically different (p<0.05, by Tukey’s test).

Wen Guo, et al. PLoS One. 2012;7(7):e40958.
2.
Figure 3

Figure 3. BME increases expression of adipogenic genes.. From: Beta-Mecaptoethanol Suppresses Inflammation and Induces Adipogenic Differentiation in 3T3-F442A Murine Preadipocytes.

A: The time course of aP2 mRNA expression in F442A cells after being induced to differentiate by insulin and fetal bovine serum and graded concentration of BME. B: Messenger RNA for SCD-1, LPL, Glut4, and adiponectin on day 10 of differentiation after being co-treated with graded concentration of BME. Results are means +/− SE (N = 3–6). Columns denoted with non-identical alphabets are statistically different (p<0.05, by Tukey’s test).

Wen Guo, et al. PLoS One. 2012;7(7):e40958.
3.
Figure 1

Figure 1. BME increases lipid accumulation in F442A cells.. From: Beta-Mecaptoethanol Suppresses Inflammation and Induces Adipogenic Differentiation in 3T3-F442A Murine Preadipocytes.

A: microphotograph of cells treated with control (left panel) and BME (1 mM, right panel) on day 6 after cells were induced to differentiated by insulin supplement. B: fluorescent microphotograph of cells after staining with BODIPY-C12 under otherwise the same conditions as those shown in A. bar = 5×10−5 m. C: quantitative fluorescent intensity of lipid staining on day 6. Results are mean +/− se from four independent cell cultures for each five different views were analyzed. Columns denoted with non-identical alphabets are statistically different (p<0.05, by Tukey’s test).

Wen Guo, et al. PLoS One. 2012;7(7):e40958.
4.
Figure 2

Figure 2. BME increases protein expression of PPARgamma and C/EBPalpha but does not directly regulate their transcription activities.. From: Beta-Mecaptoethanol Suppresses Inflammation and Induces Adipogenic Differentiation in 3T3-F442A Murine Preadipocytes.

A: representative blot of Western analysis for PPARgamma and C/EBPalpha in F442A cells differentiated without or with BME (1 mM). Cells were harvested on day 3 (d3) and day 5 (d5) after induced to differentiation by insulin and fetal bovine serum. B: quantification of PPARgamma and C/EBPalpha protein expression normalized to the loading control tubulin. Of note, although both PPARgamma1 (lower band) and PPARgamma2 (upper band) were detected, quantification was only done for PPARgamma2 because only this protein is adipocyte-specific. C: PPARgamma-driven luciferase activity in HEK293 cells treated with or without BME (1 mM) or ciglitazone (0.001 mM). Results are means +/− se (N = 3).

Wen Guo, et al. PLoS One. 2012;7(7):e40958.
5.
Figure 5

Figure 5. BME and TNFalpha mutually inhibits each other for effects on inflammation and adipogenesis.. From: Beta-Mecaptoethanol Suppresses Inflammation and Induces Adipogenic Differentiation in 3T3-F442A Murine Preadipocytes.

A: Phase-contrast microphotograph of F442A cells induced to differentiate by insulin for 9 days with or without co-treatment of BME and/or TNFalpha and indicated concentrations. B: Messenger RNA expression of selected marker genes for adipogenesis (PPARgamma, C/EBPalpha, aP2, and LPL), inflammation (iNOS and IL-6), and adipokines (adiponectin and leptin) in cells treated with TNFalpha at 0, 1 ng/ml, and 10 ng/ml without (T0, T1, T10) or with BME added at 1 mM (BT0, BT1, BT10). C. NFkappaB activity measured in HEK293 cells transfected with firefly reporter luciferase vector and control Renilla luciferase vector. Cells were treated with BME (1 mM) for 12 h before TNFalpha (10 ng/ml) was added without medium change and harvested after another 12 h of incubation (B: BME, T: TNFalpha, BT: BME plus TNFalpha). D. Western analysis for NFkappaB activation in response to TNFalpha (10 ng/ml) in differentiating preadipocytes. Cells were treated with differentiation medium with or without BME for 24 h. TNFalpha was then added directly to the culture and cells were harvested at different time points to perform Western analysis for p65ser536 and IkappaBser32 and their corresponding total protein levels, using actin as the loading index. For A–C, results are means +/− SE (N = 3–6). For B, bars denoted with non-identical alphabets are statistically different, e.g. a ≠ b, etc. (p<0.05, by Tukey’s test). For D, results are representative of three independent repeated experiments.

Wen Guo, et al. PLoS One. 2012;7(7):e40958.

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