Human/Mouse/Rat 14‑3‑3 gamma Antibody

Detection of Human, Mouse, and Rat 14‑3‑3 gamma by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 1 µg/mL Mouse Anti-Human/Mouse/Rat 14-3-3 gamma Monoclonal Antibody (Catalog # MAB5700) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for 14-3-3 gamma was detected at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human 14‑3‑3 gamma by Western Blot.

Western blot shows recombinant human 14-3-3 beta, epsilon, eta, gamma, sigma, theta, and zeta (5 ng/lane. PVDF membrane was probed with 1 µg/mL Mouse Anti-Human/Mouse/Rat 14-3-3 gamma Monoclonal Antibody (Catalog # MAB5700) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band for 14-3-3 gamma was detected at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse 14-3-3 gamma by Western Blot

Effects of adipocyte hypertrophy on Wnt5a expression. A Mature adipocytes were further cultured for 6 or 12 days & then harvested at the indicated times after induction. B Confocal microscopic images of F-actin (green), lipid vacuoles (white), & nuclei (blue) visualized by staining with Phalloidin-Alexa555, BODIPY 493/503, or Hoechst 33258, respectively (left panel). Sizes of lipid vacuoles & were measured in BODIPY stained areas & F-actin covered areas, respectively, using ImageJ. 58~74 were observed at each time point. Scale bars indicate 20 μm. C qRT-PCR analyses results of Wnt5a mRNA levels expressed as means ± SEs. All experiments were repeated independently at least twice. **p < 0.01, & ***p < 0.001 by a Student’s t-test. D–F WB of Wnt5a, beta -actin, YAP, & TAZ for whole lysates of adipocytes cultured & harvested as described in Fig. 3A. 14-3-3 gamma was used as the loading control. G WB analyses of YAP & TAZ in nuclear fraction of mature adipocytes obtained as shown Fig. 3A. Lamin A/C was used as the loading control for the nuclear fraction. H qRT-PCR of Ctgf, Ankrd1 & Cyr61 mRNA levels. I–K Mature adipocytes were treated at 18 days after adipogenic induction with Verteporfin (VP; 5 μM) for 4 h & harvested. I qRT-PCR of Wnt5a mRNA (J) WB analyses of Wnt5a protein, (K) qRT-PCR of Ctgf & Cyr61 mRNA levels (normalized versus 18 S rRNA levels). L qRT-PCR of Wnt5a, Yap, Taz, Ctgf, & Cyr61 (M) WB analyses of Wnt5a, YAP, TAZ proteins in 3T3-L1 preadipocytes transfected with control siRNA or siRNAs against YAP & TAZ. qRT-PCR results are presented as means ± SEs. Experiments were conducted independently at least twice. The significances of differences were determined using a two-tailed paired Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001; ns, not significant. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35478181), licensed under a CC-BY license. Not internally tested by R&D Systems.