human recombinant ifnλ1 (il-29) (PeproTech)
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PeproTech
human recombinant ifnλ1 (il-29)
Human Recombinant Ifnλ1 (Il 29), supplied by PeproTech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+recombinant+ifn%CE%BB1+%28il-29%29/pmc10912784-406-0-15?v=PeproTech
Average 90 stars, based on 1 article reviews
Human Recombinant Ifnλ1 (Il 29), supplied by PeproTech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+recombinant+ifn%CE%BB1+%28il-29%29/pmc10912784-406-0-15?v=PeproTech
Average 90 stars, based on 1 article reviews
human recombinant ifnλ1 (il-29) - by Bioz Stars,
2026-07
90/100 stars
Images
1) Product Images from "The population context is a driver of the heterogeneous response of epithelial cells to interferons"
Article Title: The population context is a driver of the heterogeneous response of epithelial cells to interferons
Journal: Molecular Systems Biology
doi: 10.1038/s44320-024-00011-2
Figure Legend Snippet: ( A ) Schematic depicting the T84 prom-Mx1-fp reporter cell line. Upon interaction of IFNs with their receptor, downstream signaling induces nuclear translocation of the transcription complex ISGF3. This leads to expression of the fluorescent protein under control of the ISG Mx1 promoter. The fluorescent protein accumulates in the cytosol and can be visualized by fluorescence microscopy. ( B ) Representative images showing expression of the fluorescent reporter (white) after mock, 2000 IU/mL IFNβ1, or 300 ng/mL IFNλ1-3 treatment. Nuclei are stained with DAPI (blue). n = 3 biological replicates. Scale bar = 100 µm. ( C ) T84-prom-Mx1-fp seeded at medium density were mock treated or treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3 for 24 h. The positive fluorescent cells were determined for each edge degree and the mean fluorescence intensity (MFI) was measured within each positive cell. The MFI was normalized to the mock-treatment MFI of the corresponding edge degree (normalized fluorescence). ( D ) T84-prom-Mx1-fp seeded at medium density were mock treated or treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3 for 6, 12, 24, 48, 72, and 96 h. Quantification of the percentage of total positive fluorescent cells as compared to mock-treated cells. ( E , F ) Ileum-derived organoids were seeded in 2-dimensions (2D) and treated apically with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. 24 h post-treatment, samples were fixed and indirect immunofluorescence was performed against ISG15 (green). Nuclei were stained with DAPI (blue). ( E ) Representative images are shown. Yellow arrows point at IFN responder cells located at the colony edges. Red arrows point at non-responder cells in the colony center. Scale bar = 100 µm. ( F ) Quantification of the ISG mean fluorescence intensity (arbitrary units (a.u.)) at the edge or the center of cell clusters. ( D , F ) n ≥ 3 biological replicates, error bars indicate the standard deviation. n.s. = not significant. P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by ( D ) ordinary one-way ANOVA with Dunnett’s multiple comparison test using edge degree 1 as reference and ( F ) Unpaired t test with Welch’s correction.
Techniques Used: Translocation Assay, Expressing, Control, Fluorescence, Microscopy, Staining, Derivative Assay, Immunofluorescence, Standard Deviation, Comparison
Figure Legend Snippet: ( A – E ) T84-prom-Mx1-fp seeded at medium density were mock treated or treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3 for 24 h. Cell nuclei were stained with DAPI and fluorescence microscopy was performed. ( A ) Representative images of cells (nuclei stained with DAPI are blue) expressing the fluorescent reporter (white). Yellow arrows point at IFN responder single cells or cells located at the colony edges. Red arrows point at non-responder cells in the colony center. ( B ) Correlation between single cell location and IFN-responsiveness was assessed using DBSCAN-CellX. Schematics depicting how the tool annotates cells according to their location at the edge or at the center of a cluster, or according to their edge degree are shown. ( C , D ) Quantification of the percentage of positive fluorescent cells as compared to mock-treated cells. ( C ) Edge vs. center cells. ( D ) Percentage of positive fluorescent cells dependent on the edge degree. An edge degree of 0 define single cells (no neighbors) and edge degree 1 are cells at the border of a colony. The higher the edge degree, the larger the distance from the edge. Each dot represents the percentage of positive cells dependent on the edge degree for a single well averaged over 12 individual fields of view per well. ( E ) Regression analysis and coefficient of correlation (ρ) calculated for ( D ) using a two-tailed nonparametric Spearman correlation. ( F ) T84-prom-Mx1-fp seeded at medium density were mock treated or treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3 for 6, 12, 24, 48, 72, and 96 h. Quantification of the percentage of positive fluorescent cells as compared to mock-treated cells for single, edge and center cells. ( C , D , F ) Error bars indicate standard deviations. n ≥ 3 biological replicates. n.s. = not significant. P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by ( C ) Unpaired t test with Welch’s correction, ( D ) ordinary one-way ANOVA with Dunnett’s multiple comparison test using edge degree 1 as reference and ( F ) ordinary one-way ANOVA within each time-point. .
Techniques Used: Staining, Fluorescence, Microscopy, Expressing, Two Tailed Test, Comparison
Figure Legend Snippet: ( A ) Schematic depicting the glass micropatterning approach using a Quartz-mask. ( B – D ) T84-prom-Mx1-fp cells seeded on circular micropatterns (200 µm diameter) were mock treated or treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. Fluorescent imaging was performed at 0 h, 12 h, 24 h, 48 h, 72 h, and 96 h post treatment. ( B ) Representative images at 0 h, 12 h, and 24 h post-treatment. The red line represents the edge of the patterns. Expression of the fluorescent reporter is depicted in white. Scale bar = 100 µm. ( C ) The radial distribution of immune response was determined for the 24 h post-treatment. Each population was segmented in rings (14 µm radius) and the mean fluorescence intensity (MFI) was measured within these rings. The MFI was normalized to the mock-treatment MFI of the corresponding ring. Each dot is one cell population (seeded on one micropattern). ( D ) The reporter expression for each single population was quantified by measuring the MFI at the edge and the center of a population at 12, 24, 48, 72, and 96 h post-treatment, and normalizing it to the mock-treatment of the respective time-point. Each dot is one cell population (seeded on one micropattern), lines connect edge and center of the same cell population. ( C , D ) n ≥ 3 biological replicates, in ( C ) error bars indicate the standard deviation. n.s. = not significant. P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by ( C ) RM one-was ANOVA using the edge (0–14 µm) as reference and ( D ) Paired t test. .
Techniques Used: Imaging, Expressing, Fluorescence, Standard Deviation
Figure Legend Snippet: T84 cells seeded at high and low density were mock treated, or treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. ( A ) Schematic depicting the experimental setup. ( B ) At 0, 6, 12, 24, 48, 36, and 72 h post IFN treatment, RNA was harvested to evaluate the transcription of the representative ISGs IFIT1 and Mx1 using RT-q-PCR. ISG relative expression was normalized to the mock-treated cells of the respective time-point (fold change). ( C , D ) At 0, 0.5, 1, 1.5, and 2 h post treatment, cellular protein extracts were collected to assess the phospho-STAT1 (pSTAT1) abundance by Western Blot. ( D ) For the 1 h post-treatment samples, pSTAT1 was quantified relative to the housekeeping protein α-tubulin. ( B , D ) n = 3 biological replicates, error bars indicate the standard deviation. n.s. = not significant. P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by Unpaired t test with Welch’s correction. .
Techniques Used: Expressing, Western Blot, Standard Deviation
Figure Legend Snippet: T84-prom-Mx1-fp cells at high or low density were treated with increasing concentrations of IFNβ1 and IFNλ1-3. Live cell fluorescence imaging was performed at an interval of 2 h for 24 h. ( A ) Representative images for selected time-points showing expression of the reporter prom-Mx1-mCherry (pMx1-mCh) in white. Nuclei are visualized by expression of H2B-turqiouse. Scale bar = 100 µm. ( B , C ) The mean fluorescence intensity (MFI) of the reporter expression within each cell was averaged for each density and normalized to the mock MFI of each time-point (fold change) for ( B ) IFNβ1 and ( C ) IFNλ1-3. n = 3 biological replicates. n.s = not significant, error bars indicate the standard deviation. P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by Unpaired t test with Welch’s correction between high and low density for each time-point.
Techniques Used: Fluorescence, Imaging, Expressing, Standard Deviation
Figure Legend Snippet: ( A ) Epithelial and ( B ) non-epithelial cells were seeded at high (H) and low (L) density. Cells were mock treated, or treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. 24 h post IFN treatment, RNA was harvested to evaluate the transcription of the representative ISGs IFIT1 using RT-q-PCR. ISG relative expression was normalized to the mock-treated cells (fold change). n ≥ 3 biological replicates, error bars indicate the standard deviation. n.s. = not significant. P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by Unpaired t test with Welch’s correction.
Techniques Used: Expressing, Standard Deviation
Figure Legend Snippet: ( A – D ) T84, Calu3, and human ileum-derived organoids seeded on transwell inserts were mock treated or treated from the apical (A) or basolateral (BL) side with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. ( A ) Schematic depicting the treatment of cells seeded on transwell inserts. ( B – D ) 24 h post treatment, RNA was harvested, and RT-q-PCR was used to evaluate the expression of the ISG IFIT1 and Mx1 for ( B ) T84 cells, ( C ) Calu3 cells, and ( D ) human ileum-derived organoids. Data is normalized to mock (fold change). ( E – G ) Organoids were grown in three-dimensionsional (3D) structures, either in a basolateral out (BL-out) or an apical out (A-out) conformation. ( E ) Schematic depicting BL-out and A-out organoids, and which membrane interacts with the IFNs during treatment. ( F ) BL-out and A-out organoids were stained for actin using Phalloidin (yellow), to mark the apical membrane of cells. Nuclei were stained with DAPI (blue). Scale bar = 100 µm. ( G ) BL-out and A-out organoids were treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. 24 h post treatment, RNA was harvested, and RT-q-PCR was used to evaluate the expression of the ISGs IFIT1 and Mx1. Data are normalized to mock (fold change). ( B – D , G ) n ≥ 3 biological replicates, error bars indicate the standard deviation. n.s. = not significant. P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by Unpaired t test with Welch’s correction. .
Techniques Used: Derivative Assay, Expressing, Membrane, Staining, Standard Deviation
Figure Legend Snippet: ( A – C ) T84 cells were seeded on transwell inserts to allow for a polarized monolayer formation. ( A ) 5 days post seeding, cells were fixed, and indirect immunofluorescence was performed against the junctional complex protein ZO1 (green). Nuclei were stained with DAPI (blue). Representative image is shown. Scale bars = 50 µm. n = 3 biological replicates. ( B ) Formation and integrity of the monolayer was followed by measuring the transepithelial electrical resistance (TEER) (Ω/cm 2 ) over 5 days. Values > 1000 Ω/cm 2 (dotted line) shows that cells established a polarized monolayer formation. n = 3 biological replicates. ( C ) 5 days post seeding, after reaching a polarized monolayer, the integrity of the monolayer was confirmed by the FITC-Dextran permeability assay. Diffusion of FITC-Dextran from the apical to the basolateral compartment was measured and expressed as concentration (mg/mL) of FITC-Dextran in the basolateral compartment after 3 h incubation. Positive control (pos) was the maximum diffusion possible and the negative control (neg) was medium only without FITC-Dextran. ( D ) Micropatterning of transwell inserts: Schematic depicting the micropatterning on transwell membranes using the PRIMO system (Alvéole Lab, www.alveolelab.com ). ( E , F ) T84 prom-Mx1-fp cells were seeded on micropatterned transwell membranes. Cells were mock treated, or treated simultaneously from the apical and basolateral side with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. Cells were fixed at 0 h, 12 h, and 24 h post treatment and fluorescent imaging was performed. ( E ) Representatives images showing treated T84 cell populations. The red line represents the edge of the patterns. Expression of the fluorescent reporter is depicted in white. Scale bar = 100 µm. ( F ) The reporter expression was quantified by measuring the mean fluorescence intensity (MFI) at the edge and the center of a population at 12 h or 24 h post treatment, and normalizing it to the corresponding 0 h post treatment at the edge and center, respectively. Each dot is one cell population (seeded on one micropattern), lines connect edge and center of the same cell population. ( B , C , F ) n ≥ 3 biological replicates, in ( B , C ) error bars indicate the standard deviation. n.s. = not significant, P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by ( B , C ) ordinary one-way ANOVA with Dunnett’s multiple comparison test using ( B ) day 1 or ( C ) the positive control as reference, and ( F ) Paired t test.
Techniques Used: Immunofluorescence, Staining, FITC-Dextran Permeability Assay, Diffusion-based Assay, Concentration Assay, Incubation, Positive Control, Negative Control, Imaging, Expressing, Fluorescence, Standard Deviation, Comparison
Figure Legend Snippet: ( A ) Schematic depicting paracellular diffusion in a monolayer of T84 WT and T84 ZO1 KO cells with disrupted junctional complexes. ( B ) T84 WT and T84 ZO1 KO cell protein extracts were harvested to control the absence of ZO1 protein in the KO cells by Western Blot. α-tubulin served as a housekeeping protein. Representative image is shown. ( C ) T84 WT and T84 ZO1 KO cells were fixed and indirect immunofluorescence was performed against the junctional complex protein ZO1 (green). Nuclei were stained with DAPI (blue). Representative image is shown. Scale bar = 100 µm. ( D ) T84 WT and T84 ZO1 KO cells were seeded on transwell inserts and grown as a polarized monolayer. Transepithelial electrical resistance (TEER) was measured over a period of 5 days. Dotted line shows a TEER of 1000 Ω/cm 2 corresponding to the resistance formed by confluent polarized T84 cells (Stanifer et al, ). ( E ) T84 WT and ZO1 KO cells were seeded on transwell inserts and grown as a dense monolayer, in which T84 WT cells reached a TEER > 1000 Ω/cm 2 . Cells were treated apically with 2000 IU/mL IFNβ1 and 300 ng/mL IFNλ1-3. 3 h after treatment, medium in the basolateral compartment was retrieved and IFN amount diffused from the apical to the basolateral transwell compartment was assessed by the HEK-blue assay. Depicted is the IFN concentration detected in the basolateral compartment for mock (m), IFNβ1 (β1) and IFNλ1-3 (λ) treated samples. ( F ) T84 WT and T84 ZO1 KO cells at high (H) and low (L) density were treated apically with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. 24 h post treatment, RNA was harvested, and RT-q-PCR was used to evaluate the expression of the ISG IFIT1. Data is normalized to mock (fold change). ( D – F ) n ≥ 3 biological replicates, error bars indicate the standard deviation. n.s. = not significant, P < 0.05 *, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by ( D ) multiple t tests using the Bonferroni-Dunn method and analyzing WT and ZO1 KO conditions for each time-point individually, and ( E , F ) Unpaired t test with Welch’s correction. .
Techniques Used: Diffusion-based Assay, Control, Western Blot, Immunofluorescence, Staining, Concentration Assay, Expressing, Standard Deviation
Figure Legend Snippet: T84 cells seeded at high and low density were mock-treated or pre-treated with 2000 IU/mL IFNβ1 or 300 ng/mL IFNλ1-3. 24 h post treatment, cells were infected with Vaccinia virus-eGFP (VV) or Mammalian Reovirus (MRV) at an MOI of 1 (as determined in T84 WT cells). Infection media was supplemented with the respective IFN. 16 h post infection, cells were fixed, immunostained for viral protein and fluorescence imaging analysis was performed. ( A ) Schematic of the experimental setup. ( B ) Representative images showing Vaccinia virus eGFP (green) infected cells. Nuclei were stained with DAPI (blue). Scale bar = 200 µm. ( C , D ) Quantification of the number of ( C ) Vaccinia virus eGFP infected cells and ( D ) MRV infected cells. n = 3 biological replicates, error bars indicate the standard deviation. n.s. = not significant, P < 0.01 **, P < 0.001 ***, P < 0.0001 **** as determined by ordinary one-way ANOVA with Dunnett’s multiple comparison test. Testing was performed within high or low density groups, using only virus infected cells (no pretreatment) as reference. .
Techniques Used: Infection, Virus, Fluorescence, Imaging, Staining, Standard Deviation, Comparison
