Journal: bioRxiv

Article Title: Differential miRNA Expression Contributes to Emergence of Multiple Cancer Stem Cell Subpopulations in Human Colorectal Cancer

doi: 10.1101/2023.02.06.527341

Figure Lengend Snippet: The bar graph gives the percent ALDH+, LRIG1+, CD166+, and LGR5+ cells in the HT29 CRC cell population as determined by flow cytometric analysis. Error bars represent standard error of the mean (n = 3–7 experimental replicates per marker).

Article Snippet: The following IgGs were included: mouse IgG1 – PE conjugate (5 uL for CD166; BD Biosciences #555749), purified sheep IgG (2 mg/mL for LRIG1 R&D Systems #5–001-A), and mouse IgG1-PE conjugate (10 uL for LGR5; R&D Systems #IC002P).

Techniques: Marker

Journal: bioRxiv

Article Title: Differential miRNA Expression Contributes to Emergence of Multiple Cancer Stem Cell Subpopulations in Human Colorectal Cancer

doi: 10.1101/2023.02.06.527341

Figure Lengend Snippet: This figure gives results from FACS analysis of stained CRC cells using a combination of the SC marker pair ALDH & LGR5. Panel A gives a representative dot blot graph illustrating the proportion of cells in LGR5+/ALDH−, ALDH+/LGR5−, LGR5+/ALDH+, and LGR5−/ALDH− cell subpopulations. Panel B gives a bar graph showing the average percentage positive cells in each of the four different CSC subpopulations. Error bars represent standard error of the mean. Panel C gives a heatmap from NanoString profiling analysis showing differential expression of the top 50 miRNAs in the four different isolated CSC subpopulations (increased expression = green; decreased expression = red for each CSC marker). NanoString miRNA profiling. Panel D provides the same heatmap with the respective dendrogram. Panels C and D visually illustrate the patterns that are seen when a large set of miRNAs is surveyed and they are not meant to show details, which is why the vertical axes are not legible.

Article Snippet: The following IgGs were included: mouse IgG1 – PE conjugate (5 uL for CD166; BD Biosciences #555749), purified sheep IgG (2 mg/mL for LRIG1 R&D Systems #5–001-A), and mouse IgG1-PE conjugate (10 uL for LGR5; R&D Systems #IC002P).

Techniques: Staining, Marker, Dot Blot, Expressing, Isolation

Journal: bioRxiv

Article Title: Differential miRNA Expression Contributes to Emergence of Multiple Cancer Stem Cell Subpopulations in Human Colorectal Cancer

doi: 10.1101/2023.02.06.527341

Figure Lengend Snippet: The table lists the top 10 miRNAs expressed in LGR5+/ALDH− cells. miRNAs were ranked according to p -value <0.05 for statistical significance. The asterisk depicts the original name for the miRNA that was abbreviated in the Table. The pie chart shows the functional classification identified by David analysis of the mRNAs predicted to be targeted by the miRNAs expressed in LGR5+/ALDH− cells. A summary of the functions of mRNAs predicted to be targeted by the miRNAs is given in .

Article Snippet: The following IgGs were included: mouse IgG1 – PE conjugate (5 uL for CD166; BD Biosciences #555749), purified sheep IgG (2 mg/mL for LRIG1 R&D Systems #5–001-A), and mouse IgG1-PE conjugate (10 uL for LGR5; R&D Systems #IC002P).

Techniques: Functional Assay

Journal: bioRxiv

Article Title: Differential miRNA Expression Contributes to Emergence of Multiple Cancer Stem Cell Subpopulations in Human Colorectal Cancer

doi: 10.1101/2023.02.06.527341

Figure Lengend Snippet: The table lists the top 10 miRNAs expressed in ALDH+/LGR5− cells. miRNAs were ranked according to p -value <0.05 for statistical significance. The pie chart shows the functional classification identified by David analysis of the mRNAs predicted to be targeted by the miRNAs expressed in ALDH+/LGR5− cells. A summary of the functions of mRNAs predicted to be targeted by the miRNAs is given in .

Article Snippet: The following IgGs were included: mouse IgG1 – PE conjugate (5 uL for CD166; BD Biosciences #555749), purified sheep IgG (2 mg/mL for LRIG1 R&D Systems #5–001-A), and mouse IgG1-PE conjugate (10 uL for LGR5; R&D Systems #IC002P).

Techniques: Functional Assay

Journal: bioRxiv

Article Title: Differential miRNA Expression Contributes to Emergence of Multiple Cancer Stem Cell Subpopulations in Human Colorectal Cancer

doi: 10.1101/2023.02.06.527341

Figure Lengend Snippet: The following 25 mRNAs are predicted to be targeted by miRNAs that are selectively expressed in LGR5+ cells (not expressed in ALDH+ cells): ADH1B, ADH4, ADH5, ADH6, ADH7, ALDH16A1, ALDH1A3, ALDH1L1, ALDH2, ALDH3A1, ALDH3A2, ALDH3B2, ALDH4A1, ALDH5A1, ALDH7A1, ALDH9A1, CRABP2, RBP2, RBP4, RBP5, RDH11, RDH13, RDH5, RDH8, RXRB .

Article Snippet: The following IgGs were included: mouse IgG1 – PE conjugate (5 uL for CD166; BD Biosciences #555749), purified sheep IgG (2 mg/mL for LRIG1 R&D Systems #5–001-A), and mouse IgG1-PE conjugate (10 uL for LGR5; R&D Systems #IC002P).

Techniques:

Journal: bioRxiv

Article Title: Differential miRNA Expression Contributes to Emergence of Multiple Cancer Stem Cell Subpopulations in Human Colorectal Cancer

doi: 10.1101/2023.02.06.527341

Figure Lengend Snippet: MicroRNA expression patterns in ALDH+ cells and LGR5+ cells in CSC sub-populations isolated from different FACS sorting experiments

Article Snippet: The following IgGs were included: mouse IgG1 – PE conjugate (5 uL for CD166; BD Biosciences #555749), purified sheep IgG (2 mg/mL for LRIG1 R&D Systems #5–001-A), and mouse IgG1-PE conjugate (10 uL for LGR5; R&D Systems #IC002P).

Techniques: Expressing, Isolation

Journal:

Article Title: Role of Interleukin-4 (IL-4) and IL-10 in Serum Immunoglobulin G Antibody Responses following Mucosal or Systemic Reovirus Infection

doi: 10.1128/JVI.78.7.3352-3360.2004

Figure Lengend Snippet: Reovirus-specific IgG2a (closed bars) and IgG1 (open bars) serum antibody concentrations following mucosal or parenteral infections. BALB/c mice were infected with reovirus orally, nasally, or in the FP with 3 × 106 PFU/mouse, and serum antibody responses were determined 10 days after infection. Bars indicate the standard errors among groups of three mice. The dashed line indicates the limit of detection. Data shown are from one of three similar experiments.

Article Snippet: In some instances where absolute antibody concentrations were determined, additional lanes were coated with 1 μg of goat anti-mouse Ig (H+L) (Southern Biotechnology, Birmingham, Ala.), and mouse IgG1, IgG2a, or IgA standards (Southern Biotechnology) were assayed in place of the reference serum.

Techniques: Infection

Journal:

Article Title: Role of Interleukin-4 (IL-4) and IL-10 in Serum Immunoglobulin G Antibody Responses following Mucosal or Systemic Reovirus Infection

doi: 10.1128/JVI.78.7.3352-3360.2004

Figure Lengend Snippet: Reduction of IgG2a/IgG1 ratios by rIL-4 treatment in orally infected mice. BALB/c mice were infected either orally or in the FP, and some orally infected mice received rIL-4 at 3 μg/dose (with or without 30 μg of anti-IL-4 MAb carrier/dose as described in the text) on day −1 and on day 2 of infection. The serum IgG subclass responses were determined by ELISA 10 days following infection. Data are represented as the ratio of the geometric means of IgG2a and IgG1 titers. Bars indicate the standard errors among groups of 5 to 11 mice from two independent experiments. Asterisks indicate significant differences from orally infected mice as determined by a one-way ANOVA and Dunn's post-hoc test (P < 0.05).

Article Snippet: In some instances where absolute antibody concentrations were determined, additional lanes were coated with 1 μg of goat anti-mouse Ig (H+L) (Southern Biotechnology, Birmingham, Ala.), and mouse IgG1, IgG2a, or IgA standards (Southern Biotechnology) were assayed in place of the reference serum.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Role of Interleukin-4 (IL-4) and IL-10 in Serum Immunoglobulin G Antibody Responses following Mucosal or Systemic Reovirus Infection

doi: 10.1128/JVI.78.7.3352-3360.2004

Figure Lengend Snippet: Increase in the IgG2a/IgG1 ratio in FP-infected IL-4−/− mice. IL-4−/− mice or BALB/c controls (IL-4+/+) were infected either orally or in the FP with 107 PFU of reovirus T1/L per mouse. Serum IgG subclass responses were determined by ELISA 10 days following infection. Data are represented as the ratio of the geometric mean titers of IgG2a and IgG1. Bars indicate the standard errors among groups of 4 to 10 mice from two independent experiments. The asterisk indicates a significant difference from FP-infected BALB/c control mice as determined by a one-way ANOVA and Dunn's post-hoc test (P < 0.05).

Article Snippet: In some instances where absolute antibody concentrations were determined, additional lanes were coated with 1 μg of goat anti-mouse Ig (H+L) (Southern Biotechnology, Birmingham, Ala.), and mouse IgG1, IgG2a, or IgA standards (Southern Biotechnology) were assayed in place of the reference serum.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Role of Interleukin-4 (IL-4) and IL-10 in Serum Immunoglobulin G Antibody Responses following Mucosal or Systemic Reovirus Infection

doi: 10.1128/JVI.78.7.3352-3360.2004

Figure Lengend Snippet: Partial effect of IL-10 treatment on the IgG2a/IgG1 ratio in orally infected mice. BALB/c mice were infected either orally or in the FP with 3 × 107 PFU of reovirus/mouse. Orally infected mice received rIL-10 at 100 ng/dose (or PBS as vehicle where noted) every 12 h for the first 5 days postinfection, and serum antibody responses were determined 10 days following infection. Bars indicate the standard errors among groups of four mice. Representative data are from one of three independent experiments.

Article Snippet: In some instances where absolute antibody concentrations were determined, additional lanes were coated with 1 μg of goat anti-mouse Ig (H+L) (Southern Biotechnology, Birmingham, Ala.), and mouse IgG1, IgG2a, or IgA standards (Southern Biotechnology) were assayed in place of the reference serum.

Techniques: Infection

Journal:

Article Title: Role of Interleukin-4 (IL-4) and IL-10 in Serum Immunoglobulin G Antibody Responses following Mucosal or Systemic Reovirus Infection

doi: 10.1128/JVI.78.7.3352-3360.2004

Figure Lengend Snippet: Failure of anti-IL-10R treatment to alter the IgG2a/IgG1 ratios in FP-infected mice. BALB/c mice were infected either orally or in the FP. FP-infected mice received anti-IL-10R MAb at 1.0 mg/mouse (or rat IgG1 as isotype control [IC] where noted) the day before infection and 0.5 mg/mouse every other day for 10 days. Data are represented as the ratio of the geometric means of IgG2a and IgG1 titers. Bars indicate the standard errors among groups of six to eight mice from one of two independent experiments.

Article Snippet: In some instances where absolute antibody concentrations were determined, additional lanes were coated with 1 μg of goat anti-mouse Ig (H+L) (Southern Biotechnology, Birmingham, Ala.), and mouse IgG1, IgG2a, or IgA standards (Southern Biotechnology) were assayed in place of the reference serum.

Techniques: Infection

Journal:

Article Title: Role of Interleukin-4 (IL-4) and IL-10 in Serum Immunoglobulin G Antibody Responses following Mucosal or Systemic Reovirus Infection

doi: 10.1128/JVI.78.7.3352-3360.2004

Figure Lengend Snippet: Reovirus serum IgG subclass ratio was not affected following mucosal or parenteral infections. BALB/c mice were infected with reovirus orally or in the FP with the indicated reovirus dose ranging from 104 to 109 PFU per mouse, and serum antibody responses were determined 10 days after infection. Bars indicate the standard errors among groups of four mice. Representative data are from two similar experiments.

Article Snippet: In some instances where absolute antibody concentrations were determined, additional lanes were coated with 1 μg of goat anti-mouse Ig (H+L) (Southern Biotechnology, Birmingham, Ala.), and mouse IgG1, IgG2a, or IgA standards (Southern Biotechnology) were assayed in place of the reference serum.

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Metabolic Reprograming of Cystic Fibrosis Macrophages via the IRE1α Arm of the Unfolded Protein Response Results in Exacerbated Inflammation

doi: 10.3389/fimmu.2019.01789

Figure Lengend Snippet: Table of Reagents

Article Snippet: Normal mouse IgG1 PE (Isotype) , Santa Cruz , Cat# sc-2866; RRID: AB_737219.

Techniques: Control, Purification, Recombinant, Isolation, SYBR Green Assay, Protease Inhibitor, Western Blot, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Conjugation Assay, Bicinchoninic Acid Protein Assay, Software