Author Correction: Molecular signatures written in bone proteins of 79 AD victims from Herculaneum and Pompeii

“L.B., G.N., designed research; G.N., I.R.P. and L.B. performed research; P.P. collected samples and cured the bioanthropological and taphonomic aspects of the conceptualization; G.N., I.R.P. and L.B. analysed data; F.D.P


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In the present study, the samples adopted for the bone proteomic profiling refers to human 86 remains from the 79 AD eruption victims discovered in the pyroclastic ash surge deposits at 87 Herculaneum and Pompeii 17,20 . The specimens from Herculaneum regard the victims' skeletons 88 discovered in the early '80s in a series of sea-front chambers and on the ancient beach, in the 89 suburban area of the town ( Figure S. 29). The samples belong to some of the victims found in 90 chambers 5 and 10 whose skeletons, once the appropriate surveys were carried out, were 91 removed to be studied in the laboratory, and replaced by fiberglass replicas (Figure S. 30 The skeletal elements of 15 individuals from the archaeological sites of Pompeii (7),

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Herculaneum (5) and Baia Scalandrone (3) were analysed. Table S1 shows each specimen and its 100 related information. All necessary permits were obtained for the study of the human specimen   Bone samples were prepared as described in (34) with slight modifications. Figure S2 represents 121 the whole procedure.

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Briefly, in order to prevent contamination, surfaces were removed from bone samples with  Samples were processed as reported in (89), and detailed below.

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Bone samples were eluted from the stage tips using 20 μL 40% ACN and then 10 μL 60% ACN 150 into a 96 well MS plate. Samples were concentrated under vacuum at 40ºC up to approximately 3 151 μL, and then 5 μL of 0.1% TFA and 5% ACN were added.

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Samples were then separated on a 15 cm column (75 μm inner diameter) in-house laser pulled 153 and packed with 1.9 μm C18 beads (Dr. Maisch, Germany) on an EASY-Nlc 1000 (Proxeon,154 Odense, Denmark) connected to a Q-Exactive HF (Thermo Scientific, Bremen, Germany) on a 155 77 min gradient. Buffer A was milliQ water. The peptides were separated with increasing buffer 156 B (80% ACN and 0.1% formic acid), going from 5% to 30% in 50 min, 30% to 45% in 10 min, 157 45% to 80% in 2 min, held at 80% for 5 min before dropping back down to 5% in 5 min and held 158 for 5 min. Flow rate was 250 nL/min. The column temperature was maintained at 40°C using an 159 integrated column oven. A wash-blank method using 0.1% TFA, 5% ACN was run in between 160 each sample to hinder cross contamination.

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The Q-Exactive HF was operated in data dependent top 10 mode. Spray voltage was 2 kV, S-lens 162 RF level at 50, and heated capillary to 275°C. Full scan mass spectra were recorded at a 163 resolution of 120,000 at m/z 200 over the m/z range 350-1400 with a target value of 3e6 and a 164 maximum injection time of 25 ms. HCD-generated product ions were recorded with a maximum 165 ion injection time set to 108 ms and a target value set to 2e5 and recorded at a resolution of 166 60,000. Normalized collision energy was set at 28% and the isolation window was 1.2 m/z with 167 the dynamic exclusion set to 20 s.  Evaluation of diagenetically induced chemical modifications.              substitution, respectively. The peptide with unmodified residues was also identified.