Journal: PLoS ONE

Article Title: Interleukin 6 is increased in preclinical HNSCC models of acquired cetuximab resistance, but is not required for maintenance of resistance

doi: 10.1371/journal.pone.0227261

Figure Lengend Snippet: A) PE/CA-PJ49 parental cells were transfected with 10 nM nontargeting (nt) siRNA or one of two siRNAs targeting IL6 (siIL6 A and B). RNA was extracted 96 hours post-transfection and qPCR was conducted using the IL6 primers listed in (normalized to TBP) . n = 3. *p<0.05. B) PE/CA-PJ49 parental and Ctx R cells were plated at a low density and transfected with 10 nM siRNA the next day. On the following day, and every four days thereafter, the cells were treated with vehicle (PBS) or 100 nM Ctx. The cells were stained with crystal violet 13 days post-transfection.

Article Snippet: All siRNAs were purchased from Origene (IL6 Human siRNA Oligo Duplex [SR302379], IL6R Human siRNA Oligo Duplex [SR302380], and IL6ST Human siRNA Oligo Duplex [SR302381]).

Techniques: Transfection, Staining

Journal: PLoS ONE

Article Title: Systemic siRNA Nanoparticle-Based Drugs Combined with Radiofrequency Ablation for Cancer Therapy

doi: 10.1371/journal.pone.0128910

Figure Lengend Snippet: Evaluation of distant tumor after thermal ablation without and with adjuvant MNP siRNA.

Article Snippet: Here, we show that thermal tissue ablation and nanocarrier anti-IL6 siRNA are particularly complementary as we take advantage of the fact that heating-induced vascular permeability occurs in the same geographic tissue region where infiltrating cells and increased IL-6 production is taking place, resulting in sufficiently high concentrations of anti-IL6 siRNA to render it effective in altering the cellular milieu and its composition.

Techniques: Adjuvant

Journal: The Journal of Infectious Diseases

Article Title: Role of Interleukin 6 in Innate Immunity to Mycobacterium tuberculosis Infection

doi: 10.1093/infdis/jit037

Figure Lengend Snippet: Interleukin 6 (Il6) gene silencing treatment of C57BL6/J macrophages infected with wild-type Mycobacterium tuberculosis or with Mtb:Δ-sigH, an attenuated mutant. A, Wild type–infected cells were incubated in the presence of Il6 small interfering RNA (siRNA) or a scrambled siRNA control for 24 hours. Il6 induction was evaluated by real-time polymerase chain reaction analysis performed as described in Materials and Methods. B, Culture supernatants from cells infected with the wild-type strain were assayed for Il6 by enzyme-linked immunosorbent assay. C, Tlr2−/− cells were incubated in the presence of Il6 siRNA or negative scrambled siRNA control for 24 hours. Data means ± standard errors of 3 independent experiments. *P < .05; **P < .01. Abbreviation: NS, not significant.

Article Snippet: Murine Cell Line and Il-6 siRNA Macrophage cell lines derived from C57BL6/J wild-type and Toll-like receptor 2 (TLR2)–knockout mice were obtained from BEI Resources (catalog nos.

Techniques: Infection, Mutagenesis, Incubation, Small Interfering RNA, Control, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Infectious Diseases

Article Title: Role of Interleukin 6 in Innate Immunity to Mycobacterium tuberculosis Infection

doi: 10.1093/infdis/jit037

Figure Lengend Snippet: Bacterial burden following treatment with interleukin 6 (Il6) small interfering RNA (siRNA) in C57BL6/J cells infected with wild-type Mycobacterium tuberculosis or Mtb:Δ-sigH, an attenuated mutant. Administration of Il6 siRNA for 24 hours had no significant effect on mycobacterial growth for either M. tuberculosis strain. After 48 hours of silencing, colony-forming units (CFUs) of both silenced M. tuberculosis strains were found to be significantly increased. *P < .05; **P < .01. Data means ± standard errors of 3 biological replicates.

Article Snippet: Murine Cell Line and Il-6 siRNA Macrophage cell lines derived from C57BL6/J wild-type and Toll-like receptor 2 (TLR2)–knockout mice were obtained from BEI Resources (catalog nos.

Techniques: Small Interfering RNA, Infection, Mutagenesis

Journal: The Journal of Infectious Diseases

Article Title: Role of Interleukin 6 in Innate Immunity to Mycobacterium tuberculosis Infection

doi: 10.1093/infdis/jit037

Figure Lengend Snippet: Comparison of host immunity gene expression in C57BL6/J macrophages infected with wild-type Mycobacterium tuberculosis or Mtb:Δ-sigH, an attenuated mutant, and transfected with interleukin 6 (Il6) small interfering RNA (siRNA). A, Expression of Ly6 genes. B, Expression of Cish, Csf1r, Ifitm6, Cxcl2 genes. Expression values are shown for DNA microarray experiments. Values represent responses that were statistically different between the Il-6 siRNA vs scrambled negative control treatments. *P < .05; **P < .01.

Article Snippet: Murine Cell Line and Il-6 siRNA Macrophage cell lines derived from C57BL6/J wild-type and Toll-like receptor 2 (TLR2)–knockout mice were obtained from BEI Resources (catalog nos.

Techniques: Comparison, Gene Expression, Infection, Mutagenesis, Transfection, Small Interfering RNA, Expressing, Microarray, Negative Control

Journal: The Journal of Infectious Diseases

Article Title: Role of Interleukin 6 in Innate Immunity to Mycobacterium tuberculosis Infection

doi: 10.1093/infdis/jit037

Figure Lengend Snippet: Enhanced induction of Cxcl10 by Il6 siRNA treatment. Macrophages from C57BL6/J mice were infected with wild-type Mycobacterium tuberculosis or Mtb:Δ-sigH, an attenuated mutant, and transfected with Il6 siRNA. Supernatants were collected at 24 hours after silencing and assayed for (A) Cxcl10 and (B) Ccl2 by a cytokine milliplex assay.

Article Snippet: Murine Cell Line and Il-6 siRNA Macrophage cell lines derived from C57BL6/J wild-type and Toll-like receptor 2 (TLR2)–knockout mice were obtained from BEI Resources (catalog nos.

Techniques: Infection, Mutagenesis, Transfection

Journal: bioRxiv

Article Title: The mechanism of Micafungin action to Pteropine orthoreovirus infection in human cell line

doi: 10.1101/2025.01.23.634615

Figure Lengend Snippet: IL-6 gene expression analysis with siRNA against IL-6 and MCFG. Values were calculated using the 2 -ΔΔCt comparative method. Statistical differences between groups by a student’s t-test. Bars represent the mean (n=3) relative gene expression from three independent experiments with standard deviation (*; p -value < 0.05, **; p < 0.01, ***; p < 0.001, ****; p < 0.0001) (A). Viral copy numbers by qPCR, with data represented as the mean (n=3) (B). Virus titer in the supernatant by TCID□□ assay (n=3) (C). The percentage of syncytial formation area was calculated using ImageJ software (n=12) (D). TNFSF13B gene expression analysis with siRNA against IL-6 and MCFG (E), and ICAM1 gene (F).

Article Snippet: The sequences of the siRNAs are as follows: siRNA IL-6 sense: 5’-GGACAUGACAACUCAUCUCtt-3’, siRNA IL-6 antisense: 5’-GAGAUGAGUUGUCAUGUCCtt-3’, siRNA scrambled control sense: 5’-AUUGGGUAGUGUUUCAGGCtt-3’, and siRNA scrambled control antisense: 5’-ACGUGACACGUUCGGAGAAtt-3’ (FASMAC, Kanagawa, Japan).

Techniques: Gene Expression, Standard Deviation, Virus, Software

Journal: Oncotarget

Article Title: IL-6 promotes growth and epithelial-mesenchymal transition of CD133+ cells of non-small cell lung cancer

doi: 10.18632/oncotarget.6570

Figure Lengend Snippet: ( A ) Growth analysis of CD133− cells upon exogenous IL-6 treatment. The CD133− cells of A549, H157, and H1299 cell lines were pre-treated with IL-6 (10 ng/ml) (or vehicle as control) for 5 days and cell growth at each day was analyzed by direct cell counting. ( B ) Sphere formation assay. The cell suspension containing CD133− and CD133+ cells were mixed with Matrigel (1:1, v/v) and sphere formation assays were performed. The spheres of larger than 50 μm diameter were counted. Quantification was shown on lower panel (* p < 0.05).

Article Snippet: For the incorporation of IL-6 siRNA or scrambled control plasmids into A549 and H157 cells, lentiviral plasmids carrying either control (scramble) or IL-6 siRNA (pLenti-II vector) (Applied Biological Materials Inc, Canada) sequence were transfected into 293T cells as a mixture of pLent-II-IL-6 siRNA, psPAX2 (virus-packaging plasmid), and pMD2G (envelope plasmid) (4:3:2 ratio) using PolyFect Transfection reagent (Qiagen, Valencia, CA).

Techniques: Cell Counting, Tube Formation Assay

Journal: Oncotarget

Article Title: IL-6 promotes growth and epithelial-mesenchymal transition of CD133+ cells of non-small cell lung cancer

doi: 10.18632/oncotarget.6570

Figure Lengend Snippet: ( A ) IL-6 ELISA test. IL-6 secretions in supernatants of total A549, H1299, and H157 cells were analyzed by ELISA test. The IL-6 levels secreted by 10 5 cells per 24 hours were presented. ( B and C ) qPCR results analyzing IL-6 and IL-6R levels in NSCLC cell lines. IL-6 (B) and IL-6R (C) expressions in 3 cell lines were analyzed by qPCR analyses. ( D ) qPCR analyses of IL-6 mRNA levels. The IL-6 mRNA expression in A549IL-6si/sc and H157IL-6si/sc cell sets were analyzed by qPCR tests. ( E ) Growth assay of CD133− cells obtained from A549IL-6si/A549sc and H157IL-6si/H157sc cell lines. ( F ) Sphere formation assay of CD133+ cells obtained from A549IL-6si/A549sc and H157IL-6si/H157sc cell lines. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For the incorporation of IL-6 siRNA or scrambled control plasmids into A549 and H157 cells, lentiviral plasmids carrying either control (scramble) or IL-6 siRNA (pLenti-II vector) (Applied Biological Materials Inc, Canada) sequence were transfected into 293T cells as a mixture of pLent-II-IL-6 siRNA, psPAX2 (virus-packaging plasmid), and pMD2G (envelope plasmid) (4:3:2 ratio) using PolyFect Transfection reagent (Qiagen, Valencia, CA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Growth Assay, Tube Formation Assay

Journal: Oncotarget

Article Title: IL-6 promotes growth and epithelial-mesenchymal transition of CD133+ cells of non-small cell lung cancer

doi: 10.18632/oncotarget.6570

Figure Lengend Snippet: ( A ) IF staining demonstrating IL-6 expression in CD133+ cells of A549IL-6si/sc cells used in the generation of human lung cancer xenografts. ( B ) Tumor growth in xenografts inoculated with A549IL-6si-CD133+ vs. A549sc-CD133+ cells ( n = individual tumors in each group). ( C ) Tumors excised from the xenografts derived from CD133+ cells of A549IL-6si vs. A549sc cell lines. ( D ) H & E staining (upper) and IL-6 IHC staining (lower) of tumor tissues obtained from A549IL-6si-CD133+ and A549sc-CD133+ cells inoculated mice. ( E ) Ki67 IHC staining of tumor tissues obtained from the xenografts derived from the CD133+ cells of A549IL-6si/sc cell lines (* p < 0.05).

Article Snippet: For the incorporation of IL-6 siRNA or scrambled control plasmids into A549 and H157 cells, lentiviral plasmids carrying either control (scramble) or IL-6 siRNA (pLenti-II vector) (Applied Biological Materials Inc, Canada) sequence were transfected into 293T cells as a mixture of pLent-II-IL-6 siRNA, psPAX2 (virus-packaging plasmid), and pMD2G (envelope plasmid) (4:3:2 ratio) using PolyFect Transfection reagent (Qiagen, Valencia, CA).

Techniques: Staining, Expressing, Derivative Assay, Immunohistochemistry