THP-1 Dual cells were treated with 5 µM Cyp inhibitor or DMSO for 72 hrs. After washing with PBS, cells were lysed (5 % SDS, 5 mM tris(2-carboxyethyl)phosphine (TCEP), 10 mM chloroacetamide (CAA), 100 mM Tris, 1 % Triton X-100, pH 8.5) and boiled for 10 mins followed by sonication in a water bath for 10 mins. Protein concentration was estimated by BCA assay (Thermo Fisher Scientific). Protein digestion was automated on a KingFisher APEX robot (Thermo Fisher Scientific) in 96-well format using a protocol from Koenig et al (https://star-protocols.cell.com/protocols/2975) with minor modifications. The 96-well comb is stored in plate #1, the sample in plate #2 in a final concentration of 70 % acetonitrile (ACN) and with magnetic MagReSyn Hydroxyl beads (ReSyn Biosciences) in a protein/bead ratio of 1:2. Washing solutions are in plates #3–5 (95 % ACN) and plates #6–7 (70 % ethanol). Plate #8 contains 300 μL digestion solution of 100 mM Tris pH 8.5 and trypsin (Promega) in an enzyme:protein ratio of 1:100. The protein aggregation was carried out in two steps of 1 min mixing at medium mixing speed, each followed by a 10 mins pause. The sequential washes were performed in 2.5 mins with slow speed, without releasing the beads from the magnet. The digestion was set to 16 hrs at 37 °C with slow speed. Protease activity was quenched by acidification with trifluoroacetic acid (TFA) to a final pH of 2, and the resulting peptide mixture was purified on OASIS HLB 96 well plate (Waters). Peptides were eluted twice with 100 µL of 50 % ACN and dried in a Savant DNA120 (Thermo Fisher Scientific).Peptides were then dissolved in 0.5 % TFA before liquid chromatography–tandem mass spectrometry (MS/MS) analysis. The mixture of tryptic peptides was analysed using an Ultimate3000 high-performance liquid chromatography system coupled online to an Eclipse mass spectrometer (Thermo Fisher Scientific). Buffer A consisted of water acidified with 0.1 % formic acid, while buffer B was 80 % ACN and 20 % water with 0.1 % formic acid. The peptides were first trapped for 1 min at 30 μL/min with 100% buffer A on a trap (0.3 mm by 5 mm with PepMap C18, 5 μm, 100 Å; Thermo Fisher Scientific); after trapping, the peptides were separated by a 50 cm analytical column (Acclaim PepMap, 3 μm; Thermo Fisher Scientific). The gradient was 7 to 35 % B in 103 min at 300 nL/min. Buffer B was then raised to 55 % in 3 min and increased to 99 % for the cleaning step. Peptides were ionized using a spray voltage of 2.1 kV and a capillary heated at 280 °C. The mass spectrometer was set to acquire full-scan MS spectra (350 to 1400 mass/charge ratio) for a maximum injection time set to Auto at a mass resolution of 60,000 and an automated gain control (AGC) target value of 100 %. For MSMS fragmentation we chose the DIA approach: AGC target value for fragment spectra was set at 200 %. 60 windows of 10 Da were used with an overlap of 1 Da (m/z range from 380 to 980). Resolution was set to 15,000 and IT to 40 ms. Normalised collision energy was set at 30 %.All raw files were analysed by DIA-NN v1.8.1 (DIA-NN: neural networks and interference correction enable deep proteome coverage in high throughput), searching against library generated automatically using Human reference proteome (UniProt) and standard settings: peptides from 7 to 30 AA, max number of missed cleavages of 1, oxidation (M) and protein-Acetylation as only variable modifications. The data files generated by DIA-NN were analysed using Perseus version 2.0.10.0 (https://www.nature.com/articles/nmeth.3901). The log2(x) intensities were normalised by subtracting the median intensities of each replicate across all samples, followed by the median intensities of each protein within replicate groups. Two sample t-tests were performed to identify proteins with statistically significant changes between conditions using stringency parameters s0 = 0.5 and FDR = 0.05.