Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Molecular weights and concentrations of recombinant proteins

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) μg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Concentration Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Purity of recombinant cytokine preparations and folding of recombinant aa2-152 IL-38. A . To assess the purity of the recombinant protein preparations used in this study, 1 µg of each sample (aa2-152 IL-38 with C-terminal His tag, aa2-152 IL-38 in storage buffer and in PBS, aa20-152 IL-38:Fc-KIH fusion protein, Fc-KIH IgG1 control, left gel; aa3-152 IL-38, middle gel; IL-36Ra, sIL-1Ra, icIL-1Ra1, right gel) were fractionated by SDS-PAGE and stained with Coomassie Blue. Based on calculated molecular weights (MW), expected sizes are: 18 kDa for aa2-152His IL-38 and icIL-1Ra1, 17 kDa for aa2-152 IL-38, aa3-152 IL-38, IL-36Ra and sIL-1Ra1, 28 kDa and 50 kDa for IL-38:Fc-KIH, 28 kDa for Fc-KIH IgG1 control. The size (kDa) of MW markers is indicated on the left. Storage buffer: 30 mM Hepes pH 7, 500mM NaCl, 5% glycerol, 1 mM β-ME. Fc ctrl: Fc-KIH IgG1. B . Folding of recombinant IL-38 aa2-152 was analyzed by circular dichroism. The observed percentage of β-strands was approximately 40%. CD, circular dichroism; Mdeg, measured ellipticity. C . X-ray diffraction crystal structure of the human IL-38 protein without the initiator methionine and with a Cys-to-Ser mutation at position 2 (PDB: 5BOW). β-strands (orange) represent 40.1% of the sequence, while the remaining regions (dark blue) are structured as coils and short α-helices

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) μg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Control, SDS Page, Staining, Circular Dichroism, Mutagenesis, Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Recombinant IL-38 proteins do not affect IL-36γ-induced IL-8 production in IL-36R HEK Blue and NHK cells. A . IL-36R HEK Blue and B . NHK cells were preincubated with recombinant human IL-38 variants (aa2-152 with C-terminal His tag; aa2-152; aa3-152; aa20-152 IL-38:Fc-KIH fusion protein) at indicated concentrations (10-3000 ng/ml, corresponding to 0.4–58.8 nM; see Table ), before stimulation with IL-36γ (0.1 ng/ml or 10 ng/ml, respectively, corresponding to 0.006 or 0.6 nM) for 24 h. IL-8 production was assessed by ELISA in culture supernatants. Recombinant human IL-36Ra (10-1000 ng/ml, corresponding to 0.6–58.8 nM) was used as a positive control to inhibit IL-36γ. Recombinant human sIL-1Ra (1-1000 ng/ml, corresponding to 0.06–58.8 nM) was used as a negative control. Results are expressed in % of the IL-8 production observed with IL-36γ alone. Each dot represents the mean of 3 technical replicates in an independent experiment. Results are shown as individual values and mean ± SEM for 3 ( A ) or 4 ( B ) independent experiments. ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. IL-36γ alone, as assessed by ANOVA, followed by Dunnett’s multiple comparisons test

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) μg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Positive Control, Negative Control

Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Recombinant IL-38 proteins do not globally affect IL-1α-induced IL-8 production in IL-36R HEK Blue and NHK cells. A . NHK and B . IL-36R HEK Blue cells were preincubated with recombinant human IL-38 variants (aa2-152 with C-terminal His tag; aa2-152; aa3-152; aa20-152 IL-38:Fc-KIH fusion protein) at indicated concentrations (10-3000 ng/ml, corresponding to 0.4–58.8 nM; see Table ), before stimulation with IL-1α (1 ng/ml, corresponding to 0.06 nM) for 24 h. IL-8 production was assessed by ELISA in culture supernatants. Recombinant human sIL-1Ra (1-100 ng/ml, corresponding to 0.06–5.9 nM) and recombinant icIL-1Ra1 (1-100 ng/ml, corresponding to 0.06–5.6 nM) were used as positive controls to inhibit IL-1α. Recombinant human IL-36Ra (10-1000 ng/ml, corresponding to 0.6–58.8 nM) was used as a negative control. Results are expressed in % of the IL-8 production observed with IL-1α alone. Each dot represents the mean of 3 technical replicates in an independent experiment. Results are shown as individual values and mean ± SEM for 5 ( A ) or 3 ( B ) independent experiments. ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. IL-1α alone, as assessed by ANOVA, followed by Dunnett’s multiple comparisons test

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) μg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Molecular weights and concentrations of recombinant proteins

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) µg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Concentration Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Purity of recombinant cytokine preparations and folding of recombinant aa2-152 IL-38. A . To assess the purity of the recombinant protein preparations used in this study, 1 µg of each sample (aa2-152 IL-38 with C-terminal His tag, aa2-152 IL-38 in storage buffer and in PBS, aa20-152 IL-38:Fc-KIH fusion protein, Fc-KIH IgG1 control, left gel; aa3-152 IL-38, middle gel; IL-36Ra, sIL-1Ra, icIL-1Ra1, right gel) were fractionated by SDS-PAGE and stained with Coomassie Blue. Based on calculated molecular weights (MW), expected sizes are: 18 kDa for aa2-152His IL-38 and icIL-1Ra1, 17 kDa for aa2-152 IL-38, aa3-152 IL-38, IL-36Ra and sIL-1Ra1, 28 kDa and 50 kDa for IL-38:Fc-KIH, 28 kDa for Fc-KIH IgG1 control. The size (kDa) of MW markers is indicated on the left. Storage buffer: 30 mM Hepes pH 7, 500mM NaCl, 5% glycerol, 1 mM β-ME. Fc ctrl: Fc-KIH IgG1. B . Folding of recombinant IL-38 aa2-152 was analyzed by circular dichroism. The observed percentage of β-strands was approximately 40%. CD, circular dichroism; Mdeg, measured ellipticity. C . X-ray diffraction crystal structure of the human IL-38 protein without the initiator methionine and with a Cys-to-Ser mutation at position 2 (PDB: 5BOW). β-strands (orange) represent 40.1% of the sequence, while the remaining regions (dark blue) are structured as coils and short α-helices

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) µg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Control, SDS Page, Staining, Circular Dichroism, Mutagenesis, Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Recombinant IL-38 proteins do not affect IL-36γ-induced IL-8 production in IL-36R HEK Blue and NHK cells. A . IL-36R HEK Blue and B . NHK cells were preincubated with recombinant human IL-38 variants (aa2-152 with C-terminal His tag; aa2-152; aa3-152; aa20-152 IL-38:Fc-KIH fusion protein) at indicated concentrations (10-3000 ng/ml, corresponding to 0.4–58.8 nM; see Table ), before stimulation with IL-36γ (0.1 ng/ml or 10 ng/ml, respectively, corresponding to 0.006 or 0.6 nM) for 24 h. IL-8 production was assessed by ELISA in culture supernatants. Recombinant human IL-36Ra (10-1000 ng/ml, corresponding to 0.6–58.8 nM) was used as a positive control to inhibit IL-36γ. Recombinant human sIL-1Ra (1-1000 ng/ml, corresponding to 0.06–58.8 nM) was used as a negative control. Results are expressed in % of the IL-8 production observed with IL-36γ alone. Each dot represents the mean of 3 technical replicates in an independent experiment. Results are shown as individual values and mean ± SEM for 3 ( A ) or 4 ( B ) independent experiments. ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. IL-36γ alone, as assessed by ANOVA, followed by Dunnett’s multiple comparisons test

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) µg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Positive Control, Negative Control

Journal: Cell Communication and Signaling : CCS

Article Title: Full-length and N-terminally truncated recombinant interleukin-38 variants are similarly inefficient in antagonizing interleukin-36 and interleukin-1 receptors

doi: 10.1186/s12964-025-02035-z

Figure Lengend Snippet: Recombinant IL-38 proteins do not globally affect IL-1α-induced IL-8 production in IL-36R HEK Blue and NHK cells. A . NHK and B . IL-36R HEK Blue cells were preincubated with recombinant human IL-38 variants (aa2-152 with C-terminal His tag; aa2-152; aa3-152; aa20-152 IL-38:Fc-KIH fusion protein) at indicated concentrations (10-3000 ng/ml, corresponding to 0.4–58.8 nM; see Table ), before stimulation with IL-1α (1 ng/ml, corresponding to 0.06 nM) for 24 h. IL-8 production was assessed by ELISA in culture supernatants. Recombinant human sIL-1Ra (1-100 ng/ml, corresponding to 0.06–5.9 nM) and recombinant icIL-1Ra1 (1-100 ng/ml, corresponding to 0.06–5.6 nM) were used as positive controls to inhibit IL-1α. Recombinant human IL-36Ra (10-1000 ng/ml, corresponding to 0.6–58.8 nM) was used as a negative control. Results are expressed in % of the IL-8 production observed with IL-1α alone. Each dot represents the mean of 3 technical replicates in an independent experiment. Results are shown as individual values and mean ± SEM for 5 ( A ) or 3 ( B ) independent experiments. ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. IL-1α alone, as assessed by ANOVA, followed by Dunnett’s multiple comparisons test

Article Snippet: Recombinant C-terminally His-tagged human aa2-152 IL-38 (cat n°AG-40 A-0191), aa20-152 IL-38:Fc-knobs-in-holes (KIH) linked human monomeric IL-38 (cat n°AG-40B-0226), and the corresponding human Fc-KIH IgG1 negative control (cat n° AG-35B-0015) were purchased from Adipogen Life Sciences (Fuellinsdorf, Switzerland) and resuspended at 100 (aa20-152 IL-38:Fc-KIH, Fc-KIH IgG1) or 500 (2-152 His-tagged) µg/ml in H 2 0, as recommended by the manufacturer.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Negative Control