Journal: Frontiers in Microbiology

Article Title: Probing Clostridium difficile Infection in Complex Human Gut Cellular Models

doi: 10.3389/fmicb.2019.00879

Figure Lengend Snippet: Production of C. difficile spores, toxins, and host responses to infection. (A) Colony counts of spores recovered after heat treatment, and total cells in the cell-associated C. difficile fraction (infected cell lysates) in the 2D epithelial model. Data shown are the mean of three independent experiments and error bars indicate SD, * p < 0.05 as determined by two-way ANOVA. (B) ELISA for C. difficile toxins A and B shows increased toxin production after extended infection. Toxins were measured from medium obtained from the apical compartment containing uninfected cell layers incubated for 24 or 48 h (Control) or cells infected with C. difficile for 3, 6, 24, or 48 h. Data shown are the mean of three independent experiments and error bars indicate SD, * p < 0.05 as determined by two-way ANOVA. Gray line represents the sensitivity of the test at 0.5 ng/ml. (C) ELISA for human IL-8 indicates increased IL-8 production at 24 and 48 h p.i. IL-8 was measured in medium obtained from the basolateral compartment containing uninfected cell layers incubated for 3, 6, 24, or 48 h (Control) or cells infected with C. difficile for 3–48 h. Gray line represents the limit of detection at 32 pg/ml. Data shown are the mean of three independent experiments and error bars indicate SD, * p < 0.05, ** p < 0.01, and *** p < 0.001, **** p < 0.0001 as determined by one-way ANOVA with Tukey’s test for multiple comparison.

Article Snippet: IL-8 production was also determined by analysis of basolateral supernatants from the VDC using a human IL-8 ELISA kit (R&D systems, Minneapolis, MN, United States) following the manufacturer’s instruction.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Incubation, Control, Comparison

Journal: Frontiers in Microbiology

Article Title: Probing Clostridium difficile Infection in Complex Human Gut Cellular Models

doi: 10.3389/fmicb.2019.00879

Figure Lengend Snippet: C. difficile spores and toxin production, and host response to infection in the 3D-VDC model. (A) Colony counts of spores and total cells in the host cell-associated C. difficile fraction (infected cell lysates). Data shown are the mean of three independent experiments and error bars indicate SD, ns, not significant as determined by two-way ANOVA. (B) Toxin A and B levels from apical compartment supernatants as determined by ELISA in the 3D model. Data shown are the mean of three independent experiments and error bars indicate SD, ns, not significant as determined by two-way ANOVA. Gray line represents the sensitivity of the test at 0.5 ng/ml. (C) Human IL-8 levels in supernatants from the basolateral compartments with uninfected cells incubated for 24 h or with cells infected with C. difficile for 3 and 24 h, as determined by ELISA. Gray line represents the limit of detection at 32 pg/ml. Data shown are the mean of three independent experiments and error bars indicate SD, ** p < 0.01, as determined by the one-way ANOVA with Tukey’s test for multiple comparison.

Article Snippet: IL-8 production was also determined by analysis of basolateral supernatants from the VDC using a human IL-8 ELISA kit (R&D systems, Minneapolis, MN, United States) following the manufacturer’s instruction.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Incubation, Comparison

Journal: Medical mycology

Article Title: A fucose specific lectin from Aspergillus flavus induced interleukin-8 expression is mediated by mitogen activated protein kinase p38.

doi: 10.1093/mmy/myw066

Figure Lengend Snippet: Figure 3. Interleukin-8 expression profile upon FFL treatment: Cells were treated with indicated concentrations of FFL for different time period. Total RNA was isolated and cDNA was synthesized as mentioned, mRNA expression level was determined by Q-PCR method. (A) Cell viability assay: Cells were treated with varying concentrations of FFL for 24 hours. Cell viability was determined as explained in methods. The untreated cells were considered as 100% viable. (B) Cell number assay: Cells were stimulated with varying concentrations of FFL for 24 hours. The cell number in untreated sample was taken as 100%. Trypan blue exclusion assay was used to count the cells. (C) Time dependent expression of IL-8 mRNA in L-132 cells. Cells without treatment were used as control. (D) Dose dependent expression of IL-8 mRNA in L-132 cells. (E) Inhibition of IL-8 mRNA expression by L-fucose: The 40 ng/ml of FFL was preincubated with different concentrations of L-fucose (0.1 mM, 0.2 mM, and 0.3 mM) for 1 hour prior treatment to the cells for indicated time. RNA was isolated and mRNA expression level was determined as explained earlier. (F, G) Expression of IL-8 mRNA in L-132, U-937, and PBMCs: Cells were treated with FFL for 6 and 12 hour. The mRNA expression was determined as explained earlier. (H) Measurements of IL-8 production by sandwich ELISA in L-132, U-937, and PBMCs: Cells were treated with 40 ng/ml of FFL for indicated time. After each time course supernatant was collected and IL-8 concentration was determined as mentioned in kit. The cells treated with 100 nM TPA for 4 hours was used as positive control and untreated cells considered as negative control. The data are presented as mean ± SD of three individual experiments that gave similar results. ∗∗P < .01, ∗∗∗P < .001 versus control. #P < .05, ##P < .01, ###P < .001 versus FFL (40 ng/ml).

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) Interleukin-8 concentration in cell culture supernatant was determined by sandwich ELISA using human IL-8 ELISA kit from Elabscience, Beijing (E-EL-H0048), following manufacturers protocol.

Techniques: Expressing, Isolation, Synthesized, Viability Assay, Trypan Blue Exclusion Assay, Control, Inhibition, Sandwich ELISA, Concentration Assay, Positive Control, Negative Control

Journal: Human reproduction (Oxford, England)

Article Title: Saturated fatty acids enhance TLR4 immune pathways in human trophoblasts.

doi: 10.1093/humrep/dev173

Figure Lengend Snippet: Figure 4 Effect of fatty acids on cytokine synthesis and release. Trophoblast cells were cultured with lipopolysaccharide (LPS), palmitic acid (PA), stearic acid(SA),palmitoleicacid(POA)andoleicacid(OA)for24 h.Thefinalconcentrationsinthemediumwere100 ng/mlforLPS,500 mMforPA,SA,POAand OA. Cells treated without or with 2% BSA for 24 h were used as negative controls (CTL). Levels of IL-6, IL-8 and TNF-a in the medium (B) and cell lysates (A) were measured by enzyme-linked immunosorbent assay . Data are shown as means + SEM, n ¼ 6 (A) and 3 (B), and 1.5 × 106/well primary cultured cells were plated in 12-well plate for each condition, *P , 0.05 versus CTL.

Article Snippet: Densitometric data of autoradiograms were quantified by Image J. IL-6, IL-8 and TNF-a concentration in cell lysates and culture medium were measured by enzyme-linked immunosorbent assay (ELISA) (R&D System, HS600B, HS800, and HSTA00D, S6050, S8000C, and STA00C, Minneapolis, USA).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Translational Oncology

Article Title: The expression of Slit2 and Robo1 increased during retinoic acid syndrome in acute promyelocytic leukemia and impacted differentiated cell migration

doi: 10.1016/j.tranon.2022.101370

Figure Lengend Snippet: The differences in Slit2, Robo1, and cytokines between RAS+ and other groups.

Article Snippet: Slit2 Quantitative ELISA kit (E-EL-H0931c, Elabscience, China), IL-8 ELISA kit (#Q8000B, R&D, USA), and IL-1β ELISA kit (#DLB50, R&D, USA) were used to measure the levels of Slit2, IL-8, and IL-1β, respectively. rhIL-8 (200–08 M, Peprotech, USA) and rhSlit2-N (150-11, Peprotech, USA) were used to perform the migration assay using transwell.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Translational Oncology

Article Title: The expression of Slit2 and Robo1 increased during retinoic acid syndrome in acute promyelocytic leukemia and impacted differentiated cell migration

doi: 10.1016/j.tranon.2022.101370

Figure Lengend Snippet: The correlation of Slit2, Robo1, and cytokines with the development of RAS in 16 patients.

Article Snippet: Slit2 Quantitative ELISA kit (E-EL-H0931c, Elabscience, China), IL-8 ELISA kit (#Q8000B, R&D, USA), and IL-1β ELISA kit (#DLB50, R&D, USA) were used to measure the levels of Slit2, IL-8, and IL-1β, respectively. rhIL-8 (200–08 M, Peprotech, USA) and rhSlit2-N (150-11, Peprotech, USA) were used to perform the migration assay using transwell.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Translational Oncology

Article Title: The expression of Slit2 and Robo1 increased during retinoic acid syndrome in acute promyelocytic leukemia and impacted differentiated cell migration

doi: 10.1016/j.tranon.2022.101370

Figure Lengend Snippet: Correlation between the expressions of Slit2, Robo1, and IL-8 and the severity of RAS.

Article Snippet: Slit2 Quantitative ELISA kit (E-EL-H0931c, Elabscience, China), IL-8 ELISA kit (#Q8000B, R&D, USA), and IL-1β ELISA kit (#DLB50, R&D, USA) were used to measure the levels of Slit2, IL-8, and IL-1β, respectively. rhIL-8 (200–08 M, Peprotech, USA) and rhSlit2-N (150-11, Peprotech, USA) were used to perform the migration assay using transwell.

Techniques: Enzyme-linked Immunosorbent Assay