rb mab 2217s cell signaling  (Cell Signaling Technology Inc)

Journal: The Journal of Neuroscience

Article Title: Hypothalamic miR-103 Protects from Hyperphagic Obesity in Mice

doi: 10.1523/JNEUROSCI.4251-13.2014

Figure Lengend Snippet: Enhanced PI3K-Akt-mTOR signaling causes obesity. A, B, Quantification (A) and representative microphotographs (B) of phospho-Ser235/236-S6 ribosomal protein (pS6RP) immunoreactivity in ARC of DicerCKO mice 6 weeks post-TAM (n = 4). C, Plasma insulin levels in the mice treated with 4 (DicerCKO4×TAM) or 10 (DicerCKO) injections of TAM and controls (n = 10, 5, and 5, respectively). D, X-gal staining of the forebrain in Rosa-LacZCaMKCreERT2–4×TAM mice. E, Phospho-Ser235/236-S6 ribosomal protein (pS6RP) immunoreactivity in ARC of DicerCKO4×TAM mice 6 weeks post-TAM. F, Weight in control and DicerCKO4×TAM females (n = 8 and 5, respectively). G, Food intake in control and DicerCKO4×TAM females (n = 9 and 5, respectively). H, I, Control and DicerCKO males were treated daily with either insulin (INS; n = 3) or rapamycin (RAPA; n = 4 and 3, respectively). Food intake (H) and weight (I) are presented. n = 5 and 4 for controls and DicerCKO mice in nontreated groups. J, K, Weight of control and PtenCKO female (J, n = 8 and 6, respectively) and male (K, n = 11 and 2, respectively) mice. Additionally, control and mutant females were treated with RAPA (n = 9 and 10, respectively). #, All PtenCKO females died between third and sixth weeks after TAM treatment. Scale bars: B, D, E, 500 μm.

Article Snippet: The following primary antibodies were used: phospho-S6 ribosomal protein (S235/236) Rb mAb (1:100; catalog #4857S; RRID: AB_2181035, Cell Signaling Technology) and anti-Cre antibody (1:3000; Kellendonk et al., 1999 ).

Techniques: Clinical Proteomics, Staining, Control, Mutagenesis

Journal: The Journal of Neuroscience

Article Title: Hypothalamic miR-103 Protects from Hyperphagic Obesity in Mice

doi: 10.1523/JNEUROSCI.4251-13.2014

Figure Lengend Snippet: Specific microRNAs attenuate obesity of DicerCKO mice. A, Representative images of phospho-Ser235/236-S6 ribosomal protein (pS6RP) staining in the ARC of DicerCKO mice 6 weeks after TAM treatment unilaterally infused by microRNA mixture or scrambled oligonucleotides. B, Quantification of pS6RP staining of ARC from controls and DicerCKO mice infused unilaterally with scrambled or microRNA mimics (n = 4, 6, and 4, respectively). C, D, Perirenal and perigonadal fat weights (n = 7). E, Hematoxylin and eosin staining of perigonadal adipose tissue. F, Dynamics of body weight of controls and DicerCKO mice infused unilaterally with scrambled, as well as DicerCKO mice infused with microRNA mimics to ARC (n = 6, 14, and 15, respectively). G, Food intake in Control-Scrambled, DicerCKO-Scrambled, and DicerCKO-microRNAs groups 4 weeks after TAM treatment (n = 5, 12, and 11, respectively). Scale bars: A, 250 μm; E, 100 μm.

Article Snippet: The following primary antibodies were used: phospho-S6 ribosomal protein (S235/236) Rb mAb (1:100; catalog #4857S; RRID: AB_2181035, Cell Signaling Technology) and anti-Cre antibody (1:3000; Kellendonk et al., 1999 ).

Techniques: Staining, Control

Journal: The Journal of Neuroscience

Article Title: Hypothalamic miR-103 Protects from Hyperphagic Obesity in Mice

doi: 10.1523/JNEUROSCI.4251-13.2014

Figure Lengend Snippet: Delivery of miR-103 into the ARC attenuates obesity in mice. A, Phospho-Ser235/236-S6 ribosomal protein (pS6RP) immunoreactivity in the ARC of 6-week-post-TAM DicerCKO females bilaterally injected at 4 weeks by miR-103 or miR-26b mimics and control animals. B, Weight in DicerCKO females at 5 weeks post-TAM, which were bilaterally injected to the ARC at 4 weeks post-TAM with miR-103 mimic or scrambled oligonucleotides, and control animals. n = 4, 4, and 8, respectively. C, D, Weight (C) and food intake (D) in DicerCKO females at 6 weeks post-TAM, which were bilaterally injected to the ARC at 4 weeks with low doses of miR-103 or miR-26b mimics or scrambled oligonucleotides, and control animals. n = 5, 4, 4, and 9, respectively. E, Analysis of mRNA expression array from the microdissected ARC of DicerCKO or control mice 6 weeks post-TAM (n = 3). Upregulated components of the insulin-PI3K-Akt-mTOR pathway are presented. F, G, TaqMan analysis of the Hprt-normalized Pik3r1 (F) or Pik3cg (G) expression in the microdissected ARC of 6-week-post-TAM DicerCKO mice bilaterally injected to the ARC at 4 weeks with low doses of miR-103-mimic or scrambled oligonucleotides, and control animals (n = 4, 4, 5). Scale bars: A, 500 μm.

Article Snippet: The following primary antibodies were used: phospho-S6 ribosomal protein (S235/236) Rb mAb (1:100; catalog #4857S; RRID: AB_2181035, Cell Signaling Technology) and anti-Cre antibody (1:3000; Kellendonk et al., 1999 ).

Techniques: Injection, Control, Expressing