t104865 leptin r d systems  (R&D Systems)

Journal: Cell reports

Article Title: Inter-organ insulin-leptin signal crosstalk from the liver enhances survival during food shortages.

doi: 10.1016/j.celrep.2023.112415

Figure Lengend Snippet: Figure 3. Hepatic IR KO decreases hypotha- lamic leptin sensitivity with increased plasma sLepR levels (A) Plasma leptin levels on post-tamoxifen injection day 2. (B) Representative images of pSTAT3 immunohis- tochemical staining of the hypothalamic arcuate nucleus of leptin-treated mice. Scale bar, 100mm. (C) Average number of pSTAT3-positive nuclei among hypothalamic arcuate nuclei of leptin- treated mice. (D) Representative images of Fos immunohisto- chemical staining of the hypothalamic arcuate nu- cleus of leptin-treated mice. Scale bar, 100mm. (E) Average number of Fos-positive nuclei among hypothalamic arcuate nuclei of leptin-treated mice. (F–H) Plasma sLepR levels on post-tamoxifen in- jection days 2 (F), 7 (G), and 56 (H). (I) Supernatant sLepR levels of primary culture he- patocytes isolated from IR-Control and iLIRKO mice. (J)Freeleptinindexonpost-tamoxifeninjectionday2. Data are presented as means ± SEM (*p < 0.05, **p < 0.01).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Perchloric acid NACALAI TESQUE Cat#26503-75 Recombinant mouse leptin protein R&D Systems Cat#498-OB 4% paraformaldehyde phosphate buffer solution FUJIFILM Wako Pure Chemical Corporation Cat#163-20145 Phosphate buffered saline powder FUJIFILM Wako Pure Chemical Corporation Cat#162-19321 Tissue-Tek O.C.T. compound Sakura Finetek Cat#4583 Sucrose NACALAI TESQUE Cat#30403-55 Tween 20 Merck Cat#P7949 Histofine SAB-PO(R) kit Nichirei Biosciences Cat#426042 3,30-diaminobenzidine tetrahydrochloride (DAB) FUJIFILM Wako Pure Chemical Corporation Cat#349-00903 T7 RNA polymerase Agilent Technologies Cat#600123 Liver perfusion medium Thermo Fisher Scientific Cat#17701038 HEPES Merck Cat#H4034 Collagenase from clostridium histolyticum Merck Cat#C5138 Dulbecco’s modified Eagle’s medium Merck Cat#D5796 Percoll Merck Cat#GE17-0891-01 Wortmannin FUJIFILM Wako Pure Chemical Corporation Cat#230-02341 Dimethyl sulfoxide Kanto chemical Cat#10378-01 Novolin R Novo Nordisk N/A Critical commercial assays RNeasy Mini Kit QIAGEN Cat#74104 RNeasy Micro Kit QIAGEN Cat#74004 ReverTra Ace qPCR RT Master Mix Toyobo Cat#Fsq-301 Bio-Rad Protein Assay Dye Reagent Concentrate Bio-Rad Cat#5000006 PierceTM ECL Plus Western Blotting Substrate Thermo Fisher Scientific Cat#32132 Glycogen Assay Kit Bio Vision Cat#K646-100 DIACOLOR , LIQUID TG-S Toyobo Cat#KTTG-112 Transaminase CII-B Test Wako FUJIFILM Wako Pure Chemical Corporation Cat#431-30901 Mouse/Rat Insulin ELISA Kit Morinaga Institute of Biological Science Cat#M1108 Mouse/Rat Leptin ELISA Kit Morinaga Institute of Biological Science Cat#M1305 Mouse Leptin R DuoSet ELISA R&D Systems Cat#DY497 RecoverAllTM Total Nucleic Acid Isolation Kit for FFPE Thermo Fisher Scientific Cat#AM1975 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat#4368814 Human Leptin R Quantikine ELISA Kit R&D Systems Cat#DOBR00 Deposited data Western blot images This paper Mendeley Data: https://doi.org/ 10.17632/mz3686btyp.1 Mircoarray data This paper Mendeley Data: https://doi.org/ 10.17632/mz3686btyp.1 Experimental models: Organisms/strains Mouse: C57BL/6J CLEA Japan N/A Mouse: IRloxP /loxP: B6.129S4(FVB)-Insrtm1Khn/J Jackson Laboratory JAX stock #006955 Mouse: LepRloxP /loxP: B6.129P2-Leprtm1Rck/J Jackson Laboratory JAX stock #008327 Mouse: SA-CreERT2+/ mice Schuler et al.31 N/A (Continued on next page) Cell Reports 42, 112415, May 30, 2023 15

Techniques: Clinical Proteomics, Injection, Staining, Isolation, Control