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The data collection and analysis of NMR was conducted as previously described [36 (link)]. A Bruker AVANCE IIIHD 700 MHz spectrometer equipped with a 5 mm quadruple resonance QCI-P cryoprobe (¹H, ¹³C, ¹⁵N, and ³¹P), an automatic tune and match system (ATM), and a SampleJet automated sample changer system with Bruker ICON-NMR software were utilized. The 2D ¹H−¹⁵N HSQC spectra collected for S. aureus cell lysates and culture media were assigned using a database of 2D 1H−15N HSQC reference spectra for known metabolites [36 (link)]. A chemical shift tolerance of 0.08 ppm for ¹H and 0.25 ppm for ¹⁵N were used to match metabolites to our reference database.

Venous fasting blood samples were collected in serum-tubes with no additives. The serum samples were stored at − 80 °C, until the time of metabolic profiling. The serum samples were slowly thawed at 4 °C. Aliquots of 150 μL were mixed with equal amounts of buffer solution and transferred to high-quality 3 mm MR tubes as described elsewhere [30 (link)].
The MR spectra were acquired using a Bruker Avance III 600 MHz/54 mm US-Plus (Bruker Biospin, Rheinstetten, Germany) operating at 600 MHz for proton (1H), equipped with a QCI cryoprobe. All spectra were recorded in an automatic fashion using a Bruker SampleJet and the ICON-NMR software (Bruker Biospin). Proton spectra were obtained at a constant temperature of 310 K (37 °C) using [1 (link)] a standard nuclear overhauser effect spectroscopy (NOESY) pulse sequence (Bruker: noesygppr1d) and [2 (link)] a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence with presaturation during the relaxation delay (Bruker: cpmgpr1d) to achieve water suppression, and to facilitate the detection of low-molecular-weight species by avoiding the large overlapped signals derived from large molecules, such as proteins and lipids. Measurement and processing were done in full automation using Bruker standard automation programs controlled by ICON-NMR (along with TopSpin). Chemical shift was calibrated to the middle of the alanine peaks at 1.50 ppm.

S2-013 PDAC cells and CAF 09-11 human PSCs were cultured in complete DMEM supplemented with 10% FBS and 25 mM U-¹³C₆ glucose (Cambridge Isotope Laboratories). U-¹³C₆ glucose-labelled CAF 09-11 cells were cultured with and without unlabelled tumour cell-conditioned medium. ¹³C₆-labelled polar metabolites secreted into the medium were extracted from the medium using 80% methanol extraction. All NMR experiments were conducted at 298 K using a Bruker AVANCE III-HD 700-MHz spectrometer equipped with a 5-mm quadruple resonance QCI-P cryoprobe (¹H, ¹³C, ¹⁵N and ³¹P) with z-axis gradients. A SampleJet automated sample changer system with Bruker ICON-NMR software was used to automate the NMR data collection. Two-dimensional (2D) ¹H-¹³C heteronuclear single quantum coherence (HSQC) spectra were collected and analysed as described previously^{55 (link)}.