Ceramic beads

Jejunal explants were lysed in 1 mL of Extract All reagent (Eurobio, Les Ulis, France) with ceramic beads (Bertin Technologies, Saint Quentin en Yvelines, France). Total RNA was extracted according to the manufacturer’s recommendations, as previously described [45 (link),46 (link)]. The RNA concentration was determined by measuring the optical density at 260 nm, and the RNA integrity was assessed using both NanoDrop spectrophotometric analysis (Labtech International, Paris, France) and Agilent capillary electrophoresis (Agilent 2100 Bioanalyzer, Agilent Technologies Inc., Santa Clara, CA, USA). The mean (±SD) RNA integrity number (RIN) of these mRNA preparations was 6.85 (±0.8).

After collection, 60 mg of tissue samples were cut in small pieces and lysed in 1 ml of Trizol reagent (Invitrogen, Cergy Pontoise, France) with ceramic beads (Bertin technologies, St Quentin en Yvelines, France). Total RNA was purified using RNeasy Mini Kit (Qiagen, Courtaboeuf, France) according to the manufacturer's recommendations. Residual genomic DNA was removed using DNase digestion with RNase-free DNase I Amp Grade (Invitrogen, Cergy Pontoise, France) following the recommended protocol. RNA concentration was measured by using a NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, USA), and the RNA integrity value (RIN) was assessed by using a 2100 Bioanalyzer (Agilent Technologies Inc., Santa-Clara, USA). All samples had a RIN above 8.

Isolated WapMyc tumor cells were lysed with non-ionic Triton X-100 cell lysis buffer (10 mM Tris, pH 7.5, 130 mM NaCl, 1% Triton X-100, 10 mM NaPi, 10 mM Nappi). The lysed cells were incubated on ice for 10 min and centrifuged at 13,400 rpm for 15 min at +4 °C. Snap-frozen tissue samples (3 × 3 mm) were placed in homogenization tubes containing ceramic beads (Bertin Technologies) and 300 µl of mammary gland lysis buffer (10 mM Tris-HCl, pH 7.4 (HusLab), 150 mM NaCl (Sigma), 1% Triton X-100 (Sigma), 1 mM EDTA (Sigma), 1 mM EGTA (Sigma), 0.5% NP-40 (Fluka), and 1 protease inhibitor coctail tablet (Roche) per 10 ml of solution) and homogenized with Precelly’s homogenizer (Bertin Technologies). After homogenization, the sample tubes were kept on ice for 30 min and then centrifuged 16,000 rpm for 30 min at +4 °C. The cultured cell lines were lysed with RIPA lysis buffer. SDS-page and western blot were done using standard protocols. Main manuscript western blots with original ladders are shown in Supplementary Figure 6.

Jejunal explants were lysed in 1 mL of Extract All reagent (Eurobio, Les Ulis, France) with ceramic beads (Bertin Technologies, St. Quentin en Yvelines, France). Total RNA was extracted according to the manufacturer’s recommendations as previously described^{63 (link), 64 (link)}. The RNA concentration was determined by measuring the optical density at 260 nm (OD260), and the RNA integrity was assessed using both NanoDrop spectrophotometric analysis (Nanodrop ND1000, Labtech International, Paris, France) and Agilent capillary electrophoresis (Agilent 2100 Bioanalyzer, Agilent Technologies Inc., Santa Clara, CA, USA). The mean ( ± SD) RNA Integrity Number (RIN) of these mRNA preparations was 6.85 ± 0.8.

The cardiac samples stored at −80 °C for whole-transcriptome and miRNA analysis were processed with a Precellys 24 homogenizer with ceramic beads (Bertin Technologies, Montigny-le-Bretonneux, France). RNA was isolated with TRIzol reagent and treated with DNase I. An Agilent Bioanalyzer RNA pico chip (Agilent Technologies Inc., Santa Clara, CA, USA) was used to evaluate the integrity of RNA, and a Qubit RNA kit (Thermo Fisher Scientific Inc., Waltham, MA, USA) was used to quantitate RNA.

Tissues from left ventricles and left lobe of liver were selected. Long bones were flushed with ice-cold phosphate-buffered saline to remove residual cells within bone marrow spaces. Bone specimens were chopped into small pieces. Whole brain tissues were placed with dorsal surfaces up on the cutting block. Razor blades were placed in fixed position to obtain coronal sections. In each section, hypothalamus was located and dissected as previously described (Heffner, Hartman & Seiden, 1980 (link)). Specimens from the kidneys were obtained after the surrounding connective tissues and renal capsules were carefully removed, they were then chopped into small pieces. Heart, liver, hypothalamus, and kidney were homogenized for RNA isolation following a protocol previously described (Tiyasatkulkovit et al., 2019 (link)). Bone tissues were homogenized using a Precellys Evolution Super Homogenizer with ceramic beads according to the manufacturer’s protocol (Bertin Instruments, Montigny-le-Bretonneux, France). Total RNAs were extracted using TRIZol reagent (Invitrogen, Carlsbad, CA, USA). Total RNA quality was assessed by measuring the absorbance at 260 and 280 nm using a NanoDrop-2000c spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and the 260/280 ratio ranged between 1.8 and 2.0.