Molecular biology reagents

RC and its γδ subunit were isolated as previously described [40 (link),51 (link)]. The monoclonal antibodies (mAbs) against RC, among them IIIG5 from mice and IC3 from rats, were generated and isolated as previously described [40 (link)]. The murine mAbs against the human α2A domain, JA202 and JA218, were a generous gift from D. Tuckwell (formerly of the University of Manchester, United Kingdom) [40 (link),42 (link)]. PCR primers were obtained from Eurofins (Eurofins Genomics, Germany) and are written in 5′-3′ direction. Restriction enzymes and molecular biology reagents were from Thermo Fisher Scientific (Germany) unless otherwise stated. Cloning products and expression vectors were validated by DNA sequencing (Eurofins Genomics).

For the preparation of culture media, the reagents used were obtained from the following companies: Reachim (Moscow, Russia), AppliChem (Darmstadt, Germany) and Panreac (Barcelona, Spain). Biochemical reagents were obtained from Sigma-Aldrich (Sigma, St. Louis, MO, USA), Serva (Heidelberg, Germany) and Fluka (Seelze, Germany), and molecular biology reagents were obtained from Thermo Fischer Scientific (Waltham, MA, USA), Zymo Research (Irvine, CA, USA), New England BioLabs (Hitchin, UK) and Biocom (Moscow, Russia).

Yeast strains were cultured in YP (1% yeast extract, 2% peptone) or synthetic complete (0.67% yeast nitrogen base without amino acids, SC-Ura drop-out mix) medium with 2% glycerol and 2% ethanol as carbon sources unless otherwise mentioned⁶⁹ . Both media were supplemented with 2% agar for preparing solid media. Yeast was grown in $30{}^{\circ }$ C with 230 rpm shaking in 20 mL volume in 100 mL flasks or in 50 mL volume in 250 mL flasks. Growth curves were recorded with Bioscreen C incubator (Oy Growth Curves Ab, Helsinki, Finland) as described before^{21 (link)}.
Chemicals were obtained from Merck KGaA (Darmstadt, Germany) and molecular biology reagents from Thermo Fisher Scientific (Waltham, MA, USA) unless otherwise mentioned. Lactose was obtained from VWR (Helsinki, Finland), and l-rhamnose and l-sorbose were obtained from Fluka (Charlotte, NC, USA). Cellotriose and cellotetraose were obtained from Carbosynth (Compton, Berkshire, UK) and $\alpha$ -sophorose from SERVA Electrophoresis GmbH (Heidelberg, Germany).

NAD⁺ (crystallised lithium salt, ca. 100%), NADH (disodium salt, grade II, 98%), ADP (disodium salt, 98%) and crystalline bovine serum albumin (fraction V) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Molecular biology reagents were obtained from Invitrogen (Thermo Fisher Scientific, Waltham, MA, USA). The nucleotide analogue AMPS-HDB was synthesized as previously described by Vollmer et al., 1994 [38 (link)] for the 5′-O-[S-(4-bromo-2,3-dioxobutyl)]-thiophosphate. The replacement of the 4-bromo group by hydroxyl-group was achieved by incubation (5 h) in 0.03 M 2-(N-morpholino)ethanesulfonic acid (MES) buffer, pH 7.0 at 25 °C as previously described [38 (link)]. The structure was verified by electrospray tandem mass spectrometry (ESI/MS/MS) as described in [51 (link)].

Tissue culture reagents were obtained from Corning Life Sciences B.V. (The Netherlands). Alpha-modified minimal essential media (α-MEM) without l-glutamine was purchased from Lonza (UK) and fetal bovine serum (FBS) was from Biosera (UK). Molecular biology reagents were purchased from Invitrogen (UK). Dexamethasone, alkaline phosphatase kits, and all other biochemical reagents were of analytical grade from Sigma (UK) unless otherwise stated.

All tissue culture reagents were purchased from Life Technologies (Paisley, UK); unless otherwise mentioned, other reagents were obtained from Sigma–Aldrich (Poole, Dorset, UK). Molecular biology reagents were purchased from Invitrogen (Paisley, UK) and Qiagen Ltd (Crawley, UK). Primers for RT‐PCR and qPCR were from MWG Biotech (Ebersberg, Germany) and Qiagen, respectively. The NPP1 antibody was obtained from Thermo Fisher Scientific (Loughborough, UK) and the β‐actin antibody from Abcam (Cambridge, UK).