EBF2 Antibody - #DF13398

Fig. 5: SOX4 is required for brown adipocyte differentiation in vitro. A Schematic drawing showed that the immortalized BAT SVF cells were established and differentiated into mature adipocytes. B, C The immortalized BAT SVF cells infected with scrambled or shSox4 lentivirus were differentiated into mature adipocytes as indicated in Fig. 5A. On day 6 of differentiation, mature brown adipocytes were subjected to Oil Red O staining (B) and triglyceride measurement (C) (n = 3). Scale bar, 100 μm. D, E The mRNA levels of BAT-selective genes (D) and WAT-selective genes (E) in differentiated BAT adipocytes generated as in (A) (n = 3). 18S was used as an invariant control. The mRNA levels in control cells were normalized to 1.0. F The protein levels of BAT enriched proteins and AGT in mature brown adipocytes. G RT-PCR analysis of mitochondrial number markers in mature brown adipocytes (n = 3). Ppib was used as an invariant control. The mRNA levels in control cells were normalized to 1.0. H–J BAT SVF cells were infected with scrambled or shSox4 lentivirus and analyzed for Oxygen consumption rate (OCR) at day 6 of differentiation. Oligomycin, FCCP, and Rotenone / Antimycin were added at the time points indicated by the arrows and OCR was showed in (H). The averaged basal and maximal respiration rates were shown in (I) and (J), respectively (n = 3). Asterisks (*) denote the level of statistical significance. *p 

Figure 2. MiR-19b-3p inhibits osteoblast differentiation. (A) RT-PCR analysis was used 443 to detect miRNA-19b-3p expression levels in BMSC-derived osteoblasts after 444 miR-19b-3p-overexpressed or knockdown by corresponding lentivirus. (B) ALP activity in 445 BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by 446 corresponding lentivirus. (C) RT-PCR was used to detect EBF2 mRNA expression levels in BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by 448 corresponding lentivirus. Western blot assay was used to detect (D and E) EBF2, (F-H) 449 RANKL and OPG protein expression levels in BMSC-derived osteoblasts after 450 miRNA-19b-3p overexpression or knockdown by corresponding lentivirus. (I) The ratio of 451 OPG to RANKL in BMSC-derived osteoblasts after miRNA-19b-3p overexpression or 452 knockdown by corresponding lentivirus (J) Alizarin red staining was used to detect cell 453 mineralization at day 21 after differentiation after miRNA-19b-3p overexpression or 454 knockdown at 100×. Data are analyzed using unpaired Student’s t-tests and presented as the 455 mean ± SD from three independent experiments. * p < 0.05; ** p < 0.01 vs. the indicated 456 group.