rabbit polyclonal anti- ido1 (Cell Signaling Technology Inc)
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Figure S1 A. (D and E) Serum levels of tryptophan (D) and kynurenine (E) in 10 health and 17 asthma participants were detected by mass spectrometry. (F) The ratio of Kyn to Trp is presented. Data are represented as mean ± SEM. (G) Correlation between Kyn/Trp and T1/T2 (IFN-γ/IL4+IL13) is shown. (H) The representative detail of Rabbit Polyclonal Anti Ido1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Impaired tryptophan metabolism by type 2 inflammation in epithelium worsening asthma"
Article Title: Impaired tryptophan metabolism by type 2 inflammation in epithelium worsening asthma
Journal: iScience
doi: 10.1016/j.isci.2024.109923
Figure S1 A. (D and E) Serum levels of tryptophan (D) and kynurenine (E) in 10 health and 17 asthma participants were detected by mass spectrometry. (F) The ratio of Kyn to Trp is presented. Data are represented as mean ± SEM. (G) Correlation between Kyn/Trp and T1/T2 (IFN-γ/IL4+IL13) is shown. (H) The representative detail of IDO1 IHC staining images in asthma and matched non-asthma lung tissue. Scale bar, 100 μm. See also Figure Legend Snippet: Downregulation of tryptophan metabolism in asthma (A–C) The levels of cytokines IFN-γ (A), IL-4 (B), and IL-13 (C) in the serum between control and asthma group. Data are represented as mean ± SEM. See also
Techniques Used: Control, Mass Spectrometry, Immunohistochemistry, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction
Figure S2 C. (C and D) The concentration of Trp (C) and Kyn (D) were detected after stimulation with different concentrations of IFN-γ. Data are represented as mean ± SEM. (E) The ratio of Kyn to Trp after stimulation with different concentrations of IFN-γ is presented. Data are represented as mean ± SEM. (F and G) The concentration of Trp (C) and Kyn (D) were detected after stimulation with T2 cytokines (IL-4, IL-5, and IL13) at the presence of IFN-γ. Data are represented as mean ± SEM. (H) The ratio of Kyn to Trp was calculated after treatment with T2 cytokines at the presence of IFN-γ. Data are represented as mean ± SEM. ∗∗ p < 0.05, ∗∗∗∗ p < 0.01, ns, not significant. See also Figure Legend Snippet: T1/2 cytokines regulated tryptophan metabolism to kynurenine in airway epithelium (A) The heatmap of relative mRNA levels (log10) of key enzymes (IDO1, IDO2, TDO2, and IL-4I1) in BEAS-2B cells treated with different cytokines (IFN-γ, TGF-β, IL-4, IL-5, and IL-13; all cytokines at 10 ng/mL) at different time points (4, 8, 12, 20, 24, 48 h) is presented. (B) Heatmap of metabolites related to tryptophan metabolism detected in BEAS-2B with or without 10 ng/mL IFN-γ for 24 h is shown. n = 4. See also
Techniques Used: Concentration Assay
Figure S4 . " title="IDO1 is the key enzyme regulated by T1/2 cytokines ..." property="contentUrl" width="100%" height="100%"/>
Figure Legend Snippet: IDO1 is the key enzyme regulated by T1/2 cytokines in airway epithelial cells (A) Relative mRNA level of IDO1 in BEAS-2B cells was measured after 12 h - treatment with IFN-γ at 0, 0.01, 0.1, 1 ng/mL, 10 ng/mL by RT-PCR. Data are represented as mean ± SEM. (B) IDO1 protein level was detected by western blot in BEAS-2B cells 24 and 48 h after treatment with IFN-γ at 0, 2.5, 5, 7.5, 10 ng/mL. (C and D) The expression of IDO1 at transcription (C) and expression (D) levels at different time points after IFN-γ stimulation at 100 pg/mL in BEAS-2B cells. Data are represented as mean ± SEM. (E and F) IDO1 mRNA levels after 12 h (E) and protein level after 24 h (F) in BEAS-2B after stimulation of 10 ng/mL IL-4, IL-5, and IL-13 at the presence of 100 pg/mL IFN-γ. Data are represented as mean ± SEM. (G) IDO1 mRNA levels in BEAS-2B (pretreated with si IDO1 or control) after stimulation of 10 ng/mL IFN-γ. Data are represented as mean ± SEM. (H) IDO1 expression was detected by RT-PCR after 12 h—treatment with or without Ruxolitinib (5 μM) in the presence of IFN-γ (10 ng/mL). Data are represented as mean ± SEM. (I) Western blots showed IDO1 and JAK-STAT pathway proteins level in BEAS-2B cells after 24 h stimulation of IFN-γ (10 ng/mL) with or without Ruxolitinib (5 μM). Rux, Ruxolitinib, JAK1/2 inhibitor. Data in A, C, E, and G are means ± SEM. One-way analysis of variance (ANOVA) was performed in A, C, E, and G. One of three experiments with similar results is presented. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. See also
Techniques Used: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Control
Figure S5 B. (B) IDO1 and CTH protein levels in BEAS-2B (pretreated with overexpression IDO1 plasmid or control). (C) CTH mRNA levels in BEAS-2B (pretreated with si IDO1 or control). Data are represented as mean ± SEM. (D) Under the stimulation of high concentration of IFN-γ (10 ng/ml), the expression of CTH was detected by RT-PCR at different time points. Data are represented as mean ± SEM. (E) The expression of CTH and IDO1 were detected by western Blot with the stimulation of different concentration of IFN-γ after 48 h. (F) Under the stimulation of IFN-γ or Kyn, the expression of CTH was detected by RT-PCR. Data are represented as mean ± SEM. (G) The expression of CTH and IDO1 were detected by western Blot after stimulation of 10 ng/mL IL-4, IL-5, IL-13 in the presence of 100 pg/mL IFN-γ, respectively. (H) The mRNA levels of Cth in lung were detected. Data are represented as mean ± SEM. (I) The expression of CTH was detected by western blot with the treatment of IFN-γ, Kyn, IDO/TDO inhibitors, and AhR antagonist. IACS: IDO1 & TDO2 inhibitor; LM10: TDO2 inhibitor; CH223191, AhR antagonist. Data are means ± SEM. ∗∗∗ p < 0.001. One of three repeated experiments with similar results is presented. See also Figure Legend Snippet: Kyn induces AhR translocation to regulate the transcription of the CTH gene (A) ChIP-RT-PCR of CYP1B 1 and CTH with the AHRE-related primer was performed. The amount of immunoprecipitated DNA in each sample was represented as a signal relative to the total amount of input chromatin, normalized to one. Data are represented as mean ± SEM. See also
Techniques Used: Translocation Assay, Reverse Transcription Polymerase Chain Reaction, Immunoprecipitation, Over Expression, Plasmid Preparation, Control, Concentration Assay, Expressing, Western Blot
Figure S7 . " title="IDO1 overexpression relieved HDM-induced allergic asthma (A) Representative pathological ..." property="contentUrl" width="100%" height="100%"/>
Figure Legend Snippet: IDO1 overexpression relieved HDM-induced allergic asthma (A) Representative pathological images of lung tissues by HE staining are shown. (B) Airway and vascular inflammations were assessed. # on the column was the comparison between the control and asthma group, and ∗ on the line was the comparison of the AAV-control and AAV- Ido1 group. Data are represented as mean ± SEM. (C) Subepithelial fibrosis by Sirius Red staining is shown. (D) Mucus secretion by PAS staining is shown. (E) The expression of CTH was detected by IHC. (F) The mRNA level of CTH in lung was measured by RT-PCR. Data are represented as mean ± SEM. #### p < 0.0001, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns, not significant. One of three repeated experiments with similar results is presented. The animal experiments included 6–7 BALB/c mice in each group. Scale bar, 100 μm. See also
Techniques Used: Over Expression, Staining, Comparison, Control, Expressing, Reverse Transcription Polymerase Chain Reaction
Figure Legend Snippet:
Techniques Used: Virus, Recombinant, Plasmid Preparation, shRNA, Software


