anti human il 1α  (InvivoGen)

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a, representation of the proteins with denaturing curves significantly altered (red) or unaltered (black) by 3,2-HPP treatment of THP1-Dual™ cells, using thermal proteome profiling (two combined experiments, proteomic coverage: 4301, NPARC test); b, protein-protein interaction network of proteins with denaturation curves altered by 3,2-HPP treatment (red) and the transcriptional regulators of 3, 2-HPP impacted genes in M3-9-M tumours (yellow); c, impact of 3,2-HPP treatment of THP1-Dual™ cells on the denaturation curve of GSDMD (two-combined experiments, NPARC test); d, NF-κB induction in THP1-Dual™ reporter cells treated with conditioned media harvested from WT or Gsdmd- KO THP1 cells treated with LPS ± HPP metabolites ± IL-1α and −1β neutralizing antibody (three combined experiments, one-way ANOVA with Bonferroni multiple comparison test); e-h, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT or Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); i-l, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT vs. Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS and NG ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); m, western blot of THP-1 cells treated with LPS ± NG ± HPPs (representative of two experiments).

Article Snippet: To neutralize IL-1 receptor signalling, the following antibodies were used: anti-human IL-1α (1 μg/mL; clone 7D4; mabg-hil1a-3; InvivoGen), anti-human IL-1β (1 μg/mL; clone 4H5; mabg-hil1b-3; InvivoGen) and IgG1 isotype control (1 μg/mL; clone T8E5; mabg1-ctrlm; InvivoGen).

Techniques: Comparison, Cell Culture, Western Blot

Journal: Infection and Immunity

Article Title: GM-CSF engages multiple signaling pathways to enhance pro-inflammatory cytokine responses in human monocytes during Legionella infection

doi: 10.1128/iai.00565-24

Figure Lengend Snippet: GM-CSF enhances inflammatory responses in human monocytes during Legionella infection. (A and B) THP-1 human monocytes or (C to E) primary human monocytes were pretreated with PBS or rGM-CSF for 30–60 min. Cells were then left uninfected (UI), infected with L.p . or treated with the TLR2 agonist Pam3CSK4. (A and C) Cells were harvested at 6 h post-infection (hpi) to measure IL1A , IL1B , and IL6 transcript levels by qPCR, or (B and D) cells and supernatants harvested at 24 h after infection to measure IL-1α, IL-1β, and IL-6 release by ELISA or ( E ) intracellular IL-1α and IL-1β protein levels by immunoblot with β-actin as loading control. Data represent the mean ± SEM of triplicate wells from at least three independent experiments. Data were analyzed by two-way ANOVA with Šidák’s multiple comparisons test; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, not significant.

Article Snippet: Samples were then probed with antibodies specific for IL-1α (R&D Systems MAB200), IL-1β (R&D Systems MAB201), IκBα (Cell Signaling 9242), phospho-IκBα (Cell Signaling 2859), Stat5 (Cell Signaling 94205S), phospho-Stat5 (Cell Signaling 9351S), Akt (Cell Signaling 9272), and phospho-Akt (Cell Signaling 9271).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Infection and Immunity

Article Title: GM-CSF engages multiple signaling pathways to enhance pro-inflammatory cytokine responses in human monocytes during Legionella infection

doi: 10.1128/iai.00565-24

Figure Lengend Snippet: Legionella -induced NF-κB signaling is required for GM-CSF-enhanced pro-inflammatory cytokine responses in human monocytes. ( A ) THP-1 human monocytes were pretreated with PBS or rGM-CSF for 1 h. Cells were harvested at 6 h after infection to perform immunoblot analysis for phospho-IκBα, total IκBα, or β-actin as loading control. Lanes from one membrane have been cropped and moved to depict the appropriate conditions. No changes were made to the original image during the editing. ( B ) THP-1 monocytes were pretreated with vehicle control or the IKK inhibitor BMS-345541 for 1 h. Cells were then treated with PBS or rGM-CSF for 1 h and left uninfected or infected with L.p . Cells were harvested at 6 h post-infection (hpi) to measure IL1A , IL1B , and IL6 transcript levels by qPCR. Data represent the mean ± SEM of triplicate wells from at least three independent experiments. Data were analyzed by two-way ANOVA with Šidák’s multiple comparisons test; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; ns, not significant.

Article Snippet: Samples were then probed with antibodies specific for IL-1α (R&D Systems MAB200), IL-1β (R&D Systems MAB201), IκBα (Cell Signaling 9242), phospho-IκBα (Cell Signaling 2859), Stat5 (Cell Signaling 94205S), phospho-Stat5 (Cell Signaling 9351S), Akt (Cell Signaling 9272), and phospho-Akt (Cell Signaling 9271).

Techniques: Infection, Western Blot, Control, Membrane

Journal: Infection and Immunity

Article Title: GM-CSF engages multiple signaling pathways to enhance pro-inflammatory cytokine responses in human monocytes during Legionella infection

doi: 10.1128/iai.00565-24

Figure Lengend Snippet: GM-CSF-dependent JAK2-STAT5 signaling is required to enhance inflammatory cytokine expression during Legionella infection. ( A ) THP-1 human monocytes were pretreated with PBS or rGM-CSF for 1 h. Cells were harvested at 6 h after infection to perform immunoblot analysis for phospho-STAT5, total STAT5, or β-actin as loading control. Lanes from one membrane have been cropped to depict the appropriate conditions, as indicated by the dashed lines. No changes were made to the original image during the editing. (B to C) THP-1 monocytes were pretreated with vehicle control, ( B ) the JAK2 inhibitor NVP-BSK805, or ( C ) the STAT5 inhibitor SH-4-54 for 1 h. Cells were then treated with PBS or rGM-CSF for 30–60 min and left uninfected or infected with L.p . Cells were harvested at 6 h post-infection (hpi) to measure IL1A , IL1B , and IL6 transcript levels by qPCR. Data represent the mean ± SEM of triplicate wells from at least two ( C ) or three ( B ) independent experiments. Data were analyzed by two-way ANOVA with Šidák’s multiple comparisons test; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, not significant.

Article Snippet: Samples were then probed with antibodies specific for IL-1α (R&D Systems MAB200), IL-1β (R&D Systems MAB201), IκBα (Cell Signaling 9242), phospho-IκBα (Cell Signaling 2859), Stat5 (Cell Signaling 94205S), phospho-Stat5 (Cell Signaling 9351S), Akt (Cell Signaling 9272), and phospho-Akt (Cell Signaling 9271).

Techniques: Expressing, Infection, Western Blot, Control, Membrane

Journal: Infection and Immunity

Article Title: GM-CSF engages multiple signaling pathways to enhance pro-inflammatory cytokine responses in human monocytes during Legionella infection

doi: 10.1128/iai.00565-24

Figure Lengend Snippet: PI-3K/Akt/mTORC1 signaling is required for GM-CSF-dependent pro-inflammatory cytokine expression in Legionella -infected human monocytes. (A to C) THP-1 monocytes were pretreated with vehicle control, ( A ) PI3-K inhibitor Ly294002, ( B ) Akt inhibitor MK-2206 or ( C ) mTORC1 inhibitor rapamycin for 1 h. Cells were then treated with PBS or rGM-CSF for 1 h followed by L.p . infection. Cells were harvested at 6 h after infection to measure IL1A , IL1B , and IL6 transcript levels by qPCR. Data represent the mean ± SEM of triplicate wells from at least three independent experiments. Data were analyzed by two-way ANOVA with Šidák’s multiple comparisons test; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, not significant.

Article Snippet: Samples were then probed with antibodies specific for IL-1α (R&D Systems MAB200), IL-1β (R&D Systems MAB201), IκBα (Cell Signaling 9242), phospho-IκBα (Cell Signaling 2859), Stat5 (Cell Signaling 94205S), phospho-Stat5 (Cell Signaling 9351S), Akt (Cell Signaling 9272), and phospho-Akt (Cell Signaling 9271).

Techniques: Expressing, Infection, Control

Journal: Infection and Immunity

Article Title: GM-CSF engages multiple signaling pathways to enhance pro-inflammatory cytokine responses in human monocytes during Legionella infection

doi: 10.1128/iai.00565-24

Figure Lengend Snippet: Glycolysis and amino acid metabolism are required for GM-CSF-enhanced pro-inflammatory cytokine responses in human monocytes during Legionella infection. (A to B) THP-1 monocytes were replated in ( A ) media containing glucose or galactose or ( B ) amino acid-sufficient (aa+) or -deficient (aa−) media. Cells were then treated with PBS or rGM-CSF for 1 h followed by L.p . infection. Cells were harvested at 6 h after infection to measure IL1A , IL1B , and IL6 transcript levels by qPCR. Data represent the mean ± SEM of triplicate wells from at least three independent experiments. Data were analyzed by two-way ANOVA with Šidák’s multiple comparisons test; ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, not significant.

Article Snippet: Samples were then probed with antibodies specific for IL-1α (R&D Systems MAB200), IL-1β (R&D Systems MAB201), IκBα (Cell Signaling 9242), phospho-IκBα (Cell Signaling 2859), Stat5 (Cell Signaling 94205S), phospho-Stat5 (Cell Signaling 9351S), Akt (Cell Signaling 9272), and phospho-Akt (Cell Signaling 9271).

Techniques: Infection

Journal: Nature Communications

Article Title: Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

doi: 10.1038/s41467-024-45339-3

Figure Lengend Snippet: Proteasomes generate: ( a ) non-spliced peptides via peptide hydrolysis, ( b – d ) spliced peptides through ligation of two non-contiguous splice-reactants either derived from the same molecule ( cis -spliced peptides, b ) or from two distinct molecules of the same protein (homologous trans -spliced peptides, c ) or two distinct proteins (heterologous trans -spliced peptides, d ). In ( b ), peptide fragment ligation can occur in forward order, i.e., following the orientation from N- to C-terminus of the parental protein (forward cis -peptide splicing) or in reverse order (reverse cis -peptide splicing). The two fragments, ligated by PCPS, are named splice-reactants, and their junction is named splice-site. The C-terminus of the first (N-terminal) splice-reactant is named P 1 , while the N-terminus of the second (C-terminal) splice-reactant is named P 1’ . The sequence segment between two splice-reactants is called intervening sequence. Arrows represent the substrate cleavage/splice sites used by proteasome catalytic Thr1. e Graphical representation of the experimental design of the study. The protein structure was adopted from .

Article Snippet: Degraded substrates were detected by incubating the membrane with its respective primary antibody overnight at 4 °C: mouse IL-1α (1: 1000 Abcam), human IL-1α (1:1000 Abcam), mouse IL-1β (1: 1000 R&D Systems), human IL-1β (1: 1000 Cell Signaling Technology) and human IL-37b (1:1000 Abcam).

Techniques: Ligation, Derivative Assay, Sequencing

Journal: Nature Communications

Article Title: Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

doi: 10.1038/s41467-024-45339-3

Figure Lengend Snippet: a Coomassie blue stained SDS-PAGE gel of α-Synuclein processed by human 20S standard proteasomes. A representative gel of the two biological replicates is shown. Both α-Synuclein and 20S standard proteasomes subunits are visible. b The fraction of intrinsically disordered regions in the protein sequences (computed via fraction of the sequence that had a IUPred3 score ≥0.4) and the long disorder segments reported as SLIDER score differ between efficiently degraded ( n = 15) and non-degraded ( n = 12) proteins ( p = 1.1·10 −5 and p = 0.009 for IUPred3 and SLIDER scores, respectively). Most of the degraded proteins had a SLIDER score >0.538 that is referred as the threshold for long disorder segments. Box limits represent the first (Q1) and third (Q3) quartiles, with horizontal lines depicting the median, and the interquartile range (IQR) ranging from Q1 to Q3. Whiskers denote the Q1/Q3±1.5·IQR. c The distribution of the long disorder segments reported as SLIDER score among proteins represented in the HLA-A ( n = 13,689) and HLA-B ( n = 13,755) immunopeptidomes differs from that of unrepresented proteins in HLA-I immunopeptidomes ( n = 4767), according to the IEDB database ( p = 5·10 −211 for HLA-A compared to unrepresented, p = 9·10 −211 for HLA-B compared to unrepresented, p = 0.13 for HLA-A compared to HLA-B). In ( b , c ) the horizontal lines represent the median in the plots. Statistically significant differences between groups are labeled with * (two-sided Wilcoxon rank sum test with continuity correction; p value < 0.05). Source data are provided as a file.

Article Snippet: Degraded substrates were detected by incubating the membrane with its respective primary antibody overnight at 4 °C: mouse IL-1α (1: 1000 Abcam), human IL-1α (1:1000 Abcam), mouse IL-1β (1: 1000 R&D Systems), human IL-1β (1: 1000 Cell Signaling Technology) and human IL-37b (1:1000 Abcam).

Techniques: Staining, SDS Page, Sequencing, Labeling

Journal: Nature Communications

Article Title: Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

doi: 10.1038/s41467-024-45339-3

Figure Lengend Snippet: a – g refers to database of peptide products generated by 20S proteasome degradation of the 15 proteins efficiently degraded. a Number of unique non-spliced and spliced peptides. b Distribution of abundance of non-spliced and spliced peptides, which is significantly larger than of spliced peptides ( p = 2·10 −218 and 3·10 −29 for cis - and homologous trans -spliced peptides). c Relative abundance of non-spliced and spliced peptides produced. Abundance of peptides refers to the 4 h digestion time point (3 h for α-Synuclein), which is representative of the kinetics (Supplementary Fig. ). d Spectral angle ( p = 3.6·10 −127 and p = 4.4·10 −6 ) Spearman correlation ( p = 1.7·10 −197 p = 1.2·10 −9 ) and iRT prediction error ( p = 9.1·10 −90 and p = 2.0·10 −9 ) between measurements and Prosit-predictions of non-spliced ( n = 17,265), cis- spliced (n = 2341) and homologous trans- spliced ( n = 87) peptides ( p values compare cis - and homologous trans -spliced to non-spliced). e Spectral angle ( p = 0.029) and Spearman correlation ( p = 0.0096) distribution between MS2 spectra of non-spliced ( n = 10) and cis- spliced ( n = 23) peptides compared to cognate synthetic peptides. f MS2 spectra of the representative cis -spliced peptide [G][QLGKNEEGAPQE] identified in α-Synuclein digestion, its synthetic peptide, and the synthetic peptide of its isobaric non-spliced peptide competitor [QLGKNEEGAPQEG]. Potential y-, b-, or a-ions matched between spectra are in green. Matched peaks of unknown origin are in black. Peaks not matched are in gray. Charge is shown by number + symbols. Ions’ loss of water or ammonia is symbolized by O and *. In the right panel, ion chromatograms (m/z range = 678.8163–678.8299) of the cis -spliced peptide in 0 and 24 h digestion, cognate synthetic peptide, and the synthetic isobaric non-spliced peptide are plotted. Lines delimit the range of the peak corresponding to the MS2 spectra in the left panel. g Spectral angle of the PSMs, assigned to spliced peptides ( n = 699), compared to alternative isobaric non-spliced peptide competitors for the same spectra ( p ≈ 0.0). e , g Box limits represent first and third quartiles (Q1, Q3), horizontal lines the median, whiskers the Q1/Q3 ± 1.5·IQR. b , d , e , g Statistically significant differences are labeled with * (two-sided Wilcoxon rank-sum test with continuity correction). Source data are provided as a file.

Article Snippet: Degraded substrates were detected by incubating the membrane with its respective primary antibody overnight at 4 °C: mouse IL-1α (1: 1000 Abcam), human IL-1α (1:1000 Abcam), mouse IL-1β (1: 1000 R&D Systems), human IL-1β (1: 1000 Cell Signaling Technology) and human IL-37b (1:1000 Abcam).

Techniques: Generated, Produced, Labeling

Journal: Nature Communications

Article Title: Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

doi: 10.1038/s41467-024-45339-3

Figure Lengend Snippet: a Size of theoretical peptide sequence space. Number of theoretically possible non-spliced and spliced peptide sequences that could be derived from the 15 efficiently digested proteins of this study and the 80 synthetic polypeptides in sequence-agnostic fashion. b Spectral angle distribution computed between experimentally measured and Prosit-predicted MS2 spectra of non-spliced and spliced peptides identified in protein and polypeptide digestions ( p = 5.4·10 −81 and p = 6·10 −11 for non-spliced and trans -spliced PSMs, respectively). Only PSMs of peptides shorter than 13 amino acids are included in the analysis because of the dependency of Prosit performance on peptide length. For polypeptide digestions, only those substrates that were measured on high-precision MS ( n = 25) are included in this panel for a fair comparison with the protein digestions measured with high-precision MS. c Relative frequency of non-spliced, cis - and homologous trans -spliced peptides among all peptide products. d Peptide length distribution of non-spliced and spliced peptides ( p = 0, 1.2·10 −4 , 1.2·10 −7 for non-spliced, cis - and trans -spliced peptides). e Splice-reactant lengths among cis - and trans- spliced peptides (N-terminal: p = 3.4·10 −9 , 5.2·10 −2 , C-terminal: p = 7.8·10 −8 , 1.7·10 −2 for cis - and trans -spliced peptides). f Intervening sequence lengths among cis -spliced peptides ( p = 0). g Efficacy of generation of non-spliced and spliced peptides, computed as number of detected peptide products over the number of all theoretically possible peptide products ( p = 0.03, 9.4·10 −10 , 4.9·10 −9 for non-spliced, cis - and trans -spliced peptides comparing polypeptide and protein generation efficacies; p = 1.3·10 −8 , 2.6·10 −8 comparing generation efficacies of non-spliced to cis - and trans -spliced peptides in the protein database). In ( a ), peptide product databases generated in the in vitro digestions of either proteins ( n = 15) or synthetic polypeptides ( n = 80) by 20S standard proteasomes were reported. In ( b , d – g ) statistically significant difference between the peptide product databases of proteins vs synthetic polypeptides are labeled with * (two-sided Wilcoxon rank sum test with continuity correction). In the violin plots, horizontal black lines represent the median. Source data are provided as a file. The protein structure in the cartoon corresponding to ( d ) was adopted from .

Article Snippet: Degraded substrates were detected by incubating the membrane with its respective primary antibody overnight at 4 °C: mouse IL-1α (1: 1000 Abcam), human IL-1α (1:1000 Abcam), mouse IL-1β (1: 1000 R&D Systems), human IL-1β (1: 1000 Cell Signaling Technology) and human IL-37b (1:1000 Abcam).

Techniques: Sequencing, Derivative Assay, Comparison, Generated, In Vitro, Labeling

Journal: Nature Communications

Article Title: Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

doi: 10.1038/s41467-024-45339-3

Figure Lengend Snippet: In ( a – c ), the results observed in vitro were compared to those derived from a simulated background database in silico, computed from the same substrate sequences. a Peptide length distribution of non-spliced and spliced peptides ( p = 0, 0, 3.2·10 −37 for non-spliced, cis -spliced and trans -spliced peptides). b Intervening sequence lengths of cis -spliced peptides ( p = 1.5·10 −22 ). c Splice-reactant lengths of cis- and trans -spliced peptides (N-terminal: p = 8.5·10 −251 , 3.7·10 −15 , C-terminal: p = 3.1·10 −109 , 9.4·10 −9 for cis - and trans -spliced peptides). d N- and C-terminal splice-reactants length of spliced peptides ( p = 3.2·10 −22 , 0.2 for cis -spliced and homologous trans -spliced peptides). In ( a – d ), statistically significant difference between pairs is labeled with * (two-sided Wilcoxon rank sum test with continuity correction) and qualitative analysis, i.e., counting the number of unique peptides produced by 20S proteasome whilst degrading the 15 proteins, was used. In the violin plots, horizontal black lines represent the median. Source data are provided as a file.

Article Snippet: Degraded substrates were detected by incubating the membrane with its respective primary antibody overnight at 4 °C: mouse IL-1α (1: 1000 Abcam), human IL-1α (1:1000 Abcam), mouse IL-1β (1: 1000 R&D Systems), human IL-1β (1: 1000 Cell Signaling Technology) and human IL-37b (1:1000 Abcam).

Techniques: In Vitro, Derivative Assay, In Silico, Sequencing, Labeling, Produced

Journal: Nature Communications

Article Title: Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

doi: 10.1038/s41467-024-45339-3

Figure Lengend Snippet: a , b Quantitative protein coverage profiles formed by non-spliced and spliced peptide products detected in in vitro digestion of the representative protein H3. In ( a ) the coverage is compared to the IUPred3 score predicting the presence of disordered protein segments. The gray dashed line represents the score = 0.4, which is used as threshold for a disordered segment. Predicted secondary structure elements are depicted as pink bars. The Spearman correlation coefficients and confidence intervals in the inlets denote (from left to right, respectively) correlation between: (i) spliced / non-spliced sequence coverage and IUPred3 score, (ii) spliced and non-spliced sequence coverages and (iii) spliced and non-spliced sequence coverage profiles across all time points of the digestion kinetics. Dots represent estimated Spearman correlation coefficients and error bars denote the confidence interval. Coverage profiles were normalized for spliced/non-spliced peptides separately. In ( b ) the protein coverage profiles at different digestion time points are shown. Coverage profiles were normalized for spliced/non-spliced peptides across all time points. c Scatter plot of all SCS-P 1 and PSP-P 1 computed for the 15 proteins upon in vitro digestion by 20S proteasomes. A dot represents a single protein substrate residue. The Pearson’s product moment correlation coefficient (PCC) is reported. d The Jenson-Shannon (JS) divergence of all non-spliced and spliced peptide sequences compared to a simulated background database is depicted. e The JS divergences between non-spliced and spliced peptide sequence motifs are shown. In ( a , b ) peptide amount was aggregated per substrate residue to yield quantitative coverage (see Methods for details). In ( c , d , e ) quantitative analysis, i.e., estimating the peptide amount, of the peptide products of the 15 efficiently digested proteins has been used. Source data are provided as a file.

Article Snippet: Degraded substrates were detected by incubating the membrane with its respective primary antibody overnight at 4 °C: mouse IL-1α (1: 1000 Abcam), human IL-1α (1:1000 Abcam), mouse IL-1β (1: 1000 R&D Systems), human IL-1β (1: 1000 Cell Signaling Technology) and human IL-37b (1:1000 Abcam).

Techniques: In Vitro, Sequencing, Residue

Journal: Nature Communications

Article Title: Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

doi: 10.1038/s41467-024-45339-3

Figure Lengend Snippet: a The frequency of residues used as P 1 position by peptide hydrolysis and splicing in the hotspot and non-hotspot regions of the 15 efficiently degraded proteins. The statistically significant difference between pairs is labeled with * (two-sided Wilcoxon rank sum exact test; p value = 1.8·10 −6 ). In the violin plots, horizontal black lines represent the median and dots the single proteins. b Amino acid frequency in hotspot and non-hotspot regions relative to the overall amino acid frequency among the 15 efficiently degraded proteins. Positive JS divergence indicates a dominance of these amino acids in hotspot regions, negative divergence indicates a dominance in non-hotspot regions. Peptide sequence motifs of non-spliced peptides and N-terminal splice-reactants either in hotspots ( c , d ) or non-hotspots ( e , f ). In JS divergence between the sequence motifs of either non-spliced peptides or N-terminal splice-reactants and the protein sequence background, inside ( c ) and outside ( e ) the hotspot regions. Positive values represent an overrepresentation in the specific region. In JS divergence between the sequence motifs of non-spliced peptides and N-terminal splice-reactants, inside ( d ) and outside ( f ) the hotspot regions. Positive values represent an overrepresentation among non-spliced peptides. g The JS divergence of the sequence motifs of either non-spliced peptides or N-terminal splice-reactants comparing hotspot vs non-hotspot regions. Positive values represent a prevalence in hotspots. Source data are provided as a file.

Article Snippet: Degraded substrates were detected by incubating the membrane with its respective primary antibody overnight at 4 °C: mouse IL-1α (1: 1000 Abcam), human IL-1α (1:1000 Abcam), mouse IL-1β (1: 1000 R&D Systems), human IL-1β (1: 1000 Cell Signaling Technology) and human IL-37b (1:1000 Abcam).

Techniques: Labeling, Sequencing