Structured Review

R&D Systems sil 6r
Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor <t>(sIL-6R)</t> expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
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1) Product Images from "Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation"

Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

Journal: Carcinogenesis

doi: 10.1093/carcin/bgr075

Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
Figure Legend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

Techniques Used: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P
Figure Legend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

Techniques Used: Blocking Assay, Expressing, MTT Assay

2) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

3) Product Images from "Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior"

Article Title: Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior

Journal: Journal of Neuroinflammation

doi: 10.1186/1742-2094-8-54

IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P
Figure Legend Snippet: IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P

Techniques Used:

4) Product Images from "Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation"

Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

Journal: Carcinogenesis

doi: 10.1093/carcin/bgr075

Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
Figure Legend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

Techniques Used: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P
Figure Legend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

Techniques Used: Blocking Assay, Expressing, MTT Assay

5) Product Images from "Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation"

Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

Journal: Carcinogenesis

doi: 10.1093/carcin/bgr075

Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
Figure Legend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

Techniques Used: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P
Figure Legend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

Techniques Used: Blocking Assay, Expressing, MTT Assay

6) Product Images from "An inhibitor of interleukin-6 trans-signalling, sgp130, contributes to impaired acute phase response in human chronic liver disease"

Article Title: An inhibitor of interleukin-6 trans-signalling, sgp130, contributes to impaired acute phase response in human chronic liver disease

Journal: Clinical and Experimental Immunology

doi: 10.1111/j.1365-2249.2009.03916.x

Acute phase response mediated by interleukin (IL)-6, assessed by fibrinogen (FBG) measurement in supernatants of 24-h cell cultures: (a) HepG2 cells stimulated with increasing doses of recombinant human IL-6 with or without soluble IL-6 receptor (sIL-6R). HepG2 cells (b) or HuH-7cells (c) stimulated with 50 ng/ml recombinant human IL-6, and increasing doses of gp130–Fc fusion protein, with 100 ng/ml sIL-6R (grey bars) or without sIL-6R (black bars). Values are means ± standard error of the mean of three to five experiments. § P = 0·05; * P
Figure Legend Snippet: Acute phase response mediated by interleukin (IL)-6, assessed by fibrinogen (FBG) measurement in supernatants of 24-h cell cultures: (a) HepG2 cells stimulated with increasing doses of recombinant human IL-6 with or without soluble IL-6 receptor (sIL-6R). HepG2 cells (b) or HuH-7cells (c) stimulated with 50 ng/ml recombinant human IL-6, and increasing doses of gp130–Fc fusion protein, with 100 ng/ml sIL-6R (grey bars) or without sIL-6R (black bars). Values are means ± standard error of the mean of three to five experiments. § P = 0·05; * P

Techniques Used: Recombinant

(a) Circulating interleukin (IL)-6, soluble IL-6 receptor (sIL-6R) and sgp130 levels in 15 healthy subjects (HS) and in 45 patients with alcoholic liver disease, comprising 15 with alcoholic steatohepatitis (ASH) including six with F0–F1 Kleiner fibrosis score and nine F2–F3 fibrosis stage, 15 with alcoholic cirrhosis and 15 with severe alcoholic hepatitis (AH). (b) In 84 HCV infected patients: Metavir fibrosis score F0: n = 9, F1: n = 17, F2: n = 32, F3: n = 10, F4: n = 16; * P
Figure Legend Snippet: (a) Circulating interleukin (IL)-6, soluble IL-6 receptor (sIL-6R) and sgp130 levels in 15 healthy subjects (HS) and in 45 patients with alcoholic liver disease, comprising 15 with alcoholic steatohepatitis (ASH) including six with F0–F1 Kleiner fibrosis score and nine F2–F3 fibrosis stage, 15 with alcoholic cirrhosis and 15 with severe alcoholic hepatitis (AH). (b) In 84 HCV infected patients: Metavir fibrosis score F0: n = 9, F1: n = 17, F2: n = 32, F3: n = 10, F4: n = 16; * P

Techniques Used: Infection

7) Product Images from "Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior"

Article Title: Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior

Journal: Journal of Neuroinflammation

doi: 10.1186/1742-2094-8-54

IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P
Figure Legend Snippet: IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P

Techniques Used:

8) Product Images from "Distinct role of gp130 activation in promoting self-renewal divisions by mitogenically stimulated murine hematopoietic stem cells"

Article Title: Distinct role of gp130 activation in promoting self-renewal divisions by mitogenically stimulated murine hematopoietic stem cells

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi:

Similar dose effects of different gp130-activating cytokines on CRU and CFC expansion. Sca-1 + lin − mouse BM cells were cultured in serum-free medium containing SF (50 ng/ml) and FL (100 ng/ml) and various gp130-activating cytokines at the following concentrations: IL-6 at 20 ng/ml, sIL-6R at 400 ng/ml, IL-11 at 100 ng/ml, H-IL-6 at 40 ng/ml when used alone, and H-IL-6 at 10 ng/ml when used in the four-factor combination. The fold increase in CRU and CFC (±SEM, n = 4–8) was calculated by dividing the number of these cells detected after 11 days by the corresponding numbers present in the number of Sca-1 + lin − cells used to initiate each culture.
Figure Legend Snippet: Similar dose effects of different gp130-activating cytokines on CRU and CFC expansion. Sca-1 + lin − mouse BM cells were cultured in serum-free medium containing SF (50 ng/ml) and FL (100 ng/ml) and various gp130-activating cytokines at the following concentrations: IL-6 at 20 ng/ml, sIL-6R at 400 ng/ml, IL-11 at 100 ng/ml, H-IL-6 at 40 ng/ml when used alone, and H-IL-6 at 10 ng/ml when used in the four-factor combination. The fold increase in CRU and CFC (±SEM, n = 4–8) was calculated by dividing the number of these cells detected after 11 days by the corresponding numbers present in the number of Sca-1 + lin − cells used to initiate each culture.

Techniques Used: Cell Culture

9) Product Images from "Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib"

Article Title: Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2019.00541

Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p
Figure Legend Snippet: Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p

Techniques Used: Inhibition

10) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

11) Product Images from "Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation"

Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

Journal: Carcinogenesis

doi: 10.1093/carcin/bgr075

Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
Figure Legend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

Techniques Used: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P
Figure Legend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

Techniques Used: Blocking Assay, Expressing, MTT Assay

12) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

13) Product Images from "IL-6 Trans–Signaling Links Inflammation with Angiogenesis in the Peritoneal Membrane"

Article Title: IL-6 Trans–Signaling Links Inflammation with Angiogenesis in the Peritoneal Membrane

Journal: Journal of the American Society of Nephrology : JASN

doi: 10.1681/ASN.2015101169

IL-6 signaling contributes to VEGF release by HPMCs during peritonitis. (A) IL-6 and sIL-6R levels in dialysates drained from either stable patients on PD after a routine 4-hour dwell ( n =20) or patients with peritonitis at first presentation ( n =11); the data are presented as box and whiskers plots, with the median, 25th and 75th percentiles, and range of data indicated. * P
Figure Legend Snippet: IL-6 signaling contributes to VEGF release by HPMCs during peritonitis. (A) IL-6 and sIL-6R levels in dialysates drained from either stable patients on PD after a routine 4-hour dwell ( n =20) or patients with peritonitis at first presentation ( n =11); the data are presented as box and whiskers plots, with the median, 25th and 75th percentiles, and range of data indicated. * P

Techniques Used:

Identification of sequences in the human VEGF promoter responsive to stimulation with the IL-6 + sIL-6R complex. Cells were transiently transfected with VEGF promoter constructs and stimulated with IL-6 and/or sIL-6R (both at 100 ng/ml) as indicated. Luciferase activity was determined as described in Concise Methods and expressed as relative light units (RLU). (A) Time effect of the IL-6 + sIL-6R complex on the full-length VEGF promoter activity. * P
Figure Legend Snippet: Identification of sequences in the human VEGF promoter responsive to stimulation with the IL-6 + sIL-6R complex. Cells were transiently transfected with VEGF promoter constructs and stimulated with IL-6 and/or sIL-6R (both at 100 ng/ml) as indicated. Luciferase activity was determined as described in Concise Methods and expressed as relative light units (RLU). (A) Time effect of the IL-6 + sIL-6R complex on the full-length VEGF promoter activity. * P

Techniques Used: Transfection, Construct, Luciferase, Activity Assay

Effect of SP4 on VEGF–mediated endothelial cell tube formation. (A) Cells were transiently transfected with either SP4 siRNA or scrambled (scramb.) siRNA, stimulated with IL-6 + sIL-6R (both at 100 ng/ml) for 24 hours, and then, assessed for VEGF secretion. * P
Figure Legend Snippet: Effect of SP4 on VEGF–mediated endothelial cell tube formation. (A) Cells were transiently transfected with either SP4 siRNA or scrambled (scramb.) siRNA, stimulated with IL-6 + sIL-6R (both at 100 ng/ml) for 24 hours, and then, assessed for VEGF secretion. * P

Techniques Used: Transfection

Identification of SP4 as a transcription factor mediating human VEGF promoter induction by the IL-6 + sIL-6R complex. (A and B) Cells were stimulated with IL-6 + sIL-6R at 100 ng/ml for 6 hours, and the nuclear fractions were obtained and analyzed with EMSA. In A, EMSA was performed with consensus oligonucleotide probes for SP1 and SP4. In B, formation of nuclear complexes with SP4 probe was assessed in the presence of either 100-fold molar excess of unlabeled VEGF DNA (resulting in a reduced shift) or SP4-specific antibody (resulting in supershift). (C) Effect of site-directed mutagenesis in the SP4 binding site within the VEGF promoter on its activity after stimulation with IL-6 + sIL-6R. Luciferase activity was expressed as relative light units (RLU). * P
Figure Legend Snippet: Identification of SP4 as a transcription factor mediating human VEGF promoter induction by the IL-6 + sIL-6R complex. (A and B) Cells were stimulated with IL-6 + sIL-6R at 100 ng/ml for 6 hours, and the nuclear fractions were obtained and analyzed with EMSA. In A, EMSA was performed with consensus oligonucleotide probes for SP1 and SP4. In B, formation of nuclear complexes with SP4 probe was assessed in the presence of either 100-fold molar excess of unlabeled VEGF DNA (resulting in a reduced shift) or SP4-specific antibody (resulting in supershift). (C) Effect of site-directed mutagenesis in the SP4 binding site within the VEGF promoter on its activity after stimulation with IL-6 + sIL-6R. Luciferase activity was expressed as relative light units (RLU). * P

Techniques Used: Mutagenesis, Binding Assay, Activity Assay, Luciferase

The role of STAT3 in SP4–mediated VEGF induction by the IL-6 + sIL-6R complex. (A and B) Kinetics of STAT3 and SP4 mRNA induction in HPMCs treated with IL-6 + sIL-6R (both at 100 ng/ml). * P
Figure Legend Snippet: The role of STAT3 in SP4–mediated VEGF induction by the IL-6 + sIL-6R complex. (A and B) Kinetics of STAT3 and SP4 mRNA induction in HPMCs treated with IL-6 + sIL-6R (both at 100 ng/ml). * P

Techniques Used:

Induction of VEGF in HPMCs by a combination of IL-6 and sIL-6R. (A) Kinetics of VEGF secretion by HPMCs treated with IL-6 (10 ng/ml) and sIL-6R (25 ng/ml) either singly or in combination. * P
Figure Legend Snippet: Induction of VEGF in HPMCs by a combination of IL-6 and sIL-6R. (A) Kinetics of VEGF secretion by HPMCs treated with IL-6 (10 ng/ml) and sIL-6R (25 ng/ml) either singly or in combination. * P

Techniques Used:

14) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

15) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

16) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

17) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

18) Product Images from "Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib"

Article Title: Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2019.00541

Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p
Figure Legend Snippet: Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p

Techniques Used: Inhibition

19) Product Images from "IL-6 Trans–Signaling Links Inflammation with Angiogenesis in the Peritoneal Membrane"

Article Title: IL-6 Trans–Signaling Links Inflammation with Angiogenesis in the Peritoneal Membrane

Journal: Journal of the American Society of Nephrology : JASN

doi: 10.1681/ASN.2015101169

IL-6 signaling contributes to VEGF release by HPMCs during peritonitis. (A) IL-6 and sIL-6R levels in dialysates drained from either stable patients on PD after a routine 4-hour dwell ( n =20) or patients with peritonitis at first presentation ( n =11); the data are presented as box and whiskers plots, with the median, 25th and 75th percentiles, and range of data indicated. * P
Figure Legend Snippet: IL-6 signaling contributes to VEGF release by HPMCs during peritonitis. (A) IL-6 and sIL-6R levels in dialysates drained from either stable patients on PD after a routine 4-hour dwell ( n =20) or patients with peritonitis at first presentation ( n =11); the data are presented as box and whiskers plots, with the median, 25th and 75th percentiles, and range of data indicated. * P

Techniques Used:

Identification of sequences in the human VEGF promoter responsive to stimulation with the IL-6 + sIL-6R complex. Cells were transiently transfected with VEGF promoter constructs and stimulated with IL-6 and/or sIL-6R (both at 100 ng/ml) as indicated. Luciferase activity was determined as described in Concise Methods and expressed as relative light units (RLU). (A) Time effect of the IL-6 + sIL-6R complex on the full-length VEGF promoter activity. * P
Figure Legend Snippet: Identification of sequences in the human VEGF promoter responsive to stimulation with the IL-6 + sIL-6R complex. Cells were transiently transfected with VEGF promoter constructs and stimulated with IL-6 and/or sIL-6R (both at 100 ng/ml) as indicated. Luciferase activity was determined as described in Concise Methods and expressed as relative light units (RLU). (A) Time effect of the IL-6 + sIL-6R complex on the full-length VEGF promoter activity. * P

Techniques Used: Transfection, Construct, Luciferase, Activity Assay

Effect of SP4 on VEGF–mediated endothelial cell tube formation. (A) Cells were transiently transfected with either SP4 siRNA or scrambled (scramb.) siRNA, stimulated with IL-6 + sIL-6R (both at 100 ng/ml) for 24 hours, and then, assessed for VEGF secretion. * P
Figure Legend Snippet: Effect of SP4 on VEGF–mediated endothelial cell tube formation. (A) Cells were transiently transfected with either SP4 siRNA or scrambled (scramb.) siRNA, stimulated with IL-6 + sIL-6R (both at 100 ng/ml) for 24 hours, and then, assessed for VEGF secretion. * P

Techniques Used: Transfection

Identification of SP4 as a transcription factor mediating human VEGF promoter induction by the IL-6 + sIL-6R complex. (A and B) Cells were stimulated with IL-6 + sIL-6R at 100 ng/ml for 6 hours, and the nuclear fractions were obtained and analyzed with EMSA. In A, EMSA was performed with consensus oligonucleotide probes for SP1 and SP4. In B, formation of nuclear complexes with SP4 probe was assessed in the presence of either 100-fold molar excess of unlabeled VEGF DNA (resulting in a reduced shift) or SP4-specific antibody (resulting in supershift). (C) Effect of site-directed mutagenesis in the SP4 binding site within the VEGF promoter on its activity after stimulation with IL-6 + sIL-6R. Luciferase activity was expressed as relative light units (RLU). * P
Figure Legend Snippet: Identification of SP4 as a transcription factor mediating human VEGF promoter induction by the IL-6 + sIL-6R complex. (A and B) Cells were stimulated with IL-6 + sIL-6R at 100 ng/ml for 6 hours, and the nuclear fractions were obtained and analyzed with EMSA. In A, EMSA was performed with consensus oligonucleotide probes for SP1 and SP4. In B, formation of nuclear complexes with SP4 probe was assessed in the presence of either 100-fold molar excess of unlabeled VEGF DNA (resulting in a reduced shift) or SP4-specific antibody (resulting in supershift). (C) Effect of site-directed mutagenesis in the SP4 binding site within the VEGF promoter on its activity after stimulation with IL-6 + sIL-6R. Luciferase activity was expressed as relative light units (RLU). * P

Techniques Used: Mutagenesis, Binding Assay, Activity Assay, Luciferase

The role of STAT3 in SP4–mediated VEGF induction by the IL-6 + sIL-6R complex. (A and B) Kinetics of STAT3 and SP4 mRNA induction in HPMCs treated with IL-6 + sIL-6R (both at 100 ng/ml). * P
Figure Legend Snippet: The role of STAT3 in SP4–mediated VEGF induction by the IL-6 + sIL-6R complex. (A and B) Kinetics of STAT3 and SP4 mRNA induction in HPMCs treated with IL-6 + sIL-6R (both at 100 ng/ml). * P

Techniques Used:

Induction of VEGF in HPMCs by a combination of IL-6 and sIL-6R. (A) Kinetics of VEGF secretion by HPMCs treated with IL-6 (10 ng/ml) and sIL-6R (25 ng/ml) either singly or in combination. * P
Figure Legend Snippet: Induction of VEGF in HPMCs by a combination of IL-6 and sIL-6R. (A) Kinetics of VEGF secretion by HPMCs treated with IL-6 (10 ng/ml) and sIL-6R (25 ng/ml) either singly or in combination. * P

Techniques Used:

20) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

21) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

22) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

23) Product Images from "Distinct role of gp130 activation in promoting self-renewal divisions by mitogenically stimulated murine hematopoietic stem cells"

Article Title: Distinct role of gp130 activation in promoting self-renewal divisions by mitogenically stimulated murine hematopoietic stem cells

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi:

Similar dose effects of different gp130-activating cytokines on CRU and CFC expansion. Sca-1 + lin − mouse BM cells were cultured in serum-free medium containing SF (50 ng/ml) and FL (100 ng/ml) and various gp130-activating cytokines at the following concentrations: IL-6 at 20 ng/ml, sIL-6R at 400 ng/ml, IL-11 at 100 ng/ml, H-IL-6 at 40 ng/ml when used alone, and H-IL-6 at 10 ng/ml when used in the four-factor combination. The fold increase in CRU and CFC (±SEM, n = 4–8) was calculated by dividing the number of these cells detected after 11 days by the corresponding numbers present in the number of Sca-1 + lin − cells used to initiate each culture.
Figure Legend Snippet: Similar dose effects of different gp130-activating cytokines on CRU and CFC expansion. Sca-1 + lin − mouse BM cells were cultured in serum-free medium containing SF (50 ng/ml) and FL (100 ng/ml) and various gp130-activating cytokines at the following concentrations: IL-6 at 20 ng/ml, sIL-6R at 400 ng/ml, IL-11 at 100 ng/ml, H-IL-6 at 40 ng/ml when used alone, and H-IL-6 at 10 ng/ml when used in the four-factor combination. The fold increase in CRU and CFC (±SEM, n = 4–8) was calculated by dividing the number of these cells detected after 11 days by the corresponding numbers present in the number of Sca-1 + lin − cells used to initiate each culture.

Techniques Used: Cell Culture

24) Product Images from "Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation"

Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

Journal: Carcinogenesis

doi: 10.1093/carcin/bgr075

Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
Figure Legend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

Techniques Used: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P
Figure Legend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

Techniques Used: Blocking Assay, Expressing, MTT Assay

25) Product Images from "Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation"

Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

Journal: Carcinogenesis

doi: 10.1093/carcin/bgr075

Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
Figure Legend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

Techniques Used: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P
Figure Legend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

Techniques Used: Blocking Assay, Expressing, MTT Assay

26) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

27) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

28) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

29) Product Images from "CRITICAL ROLE OF STAT3 IN IL-6-MEDIATED DRUG RESISTANCE IN HUMAN NEUROBLASTOMA"

Article Title: CRITICAL ROLE OF STAT3 IN IL-6-MEDIATED DRUG RESISTANCE IN HUMAN NEUROBLASTOMA

Journal: Cancer research

doi: 10.1158/0008-5472.CAN-12-2353

IL-6 and sIL-6R protect neuroblastoma cells from drug toxicity. CHLA-255 (A and B) and SK-N-SH cells (C and D) were treated with IL-6 (10 ng/mL) alone or with sIL-6R (25 ng/mL) for 24 hours before being exposed to etoposide (A and C) or melphalan (B and
Figure Legend Snippet: IL-6 and sIL-6R protect neuroblastoma cells from drug toxicity. CHLA-255 (A and B) and SK-N-SH cells (C and D) were treated with IL-6 (10 ng/mL) alone or with sIL-6R (25 ng/mL) for 24 hours before being exposed to etoposide (A and C) or melphalan (B and

Techniques Used:

IL-6 and sIL-6R protect neuroblastoma cells from drug-induced apoptosis. A and B, CHLA-255 cells were treated as in . After 24 hours mitochondrial membrane depolarization (MΨP) was examined by JC-1 staining by flow cytometry. The data
Figure Legend Snippet: IL-6 and sIL-6R protect neuroblastoma cells from drug-induced apoptosis. A and B, CHLA-255 cells were treated as in . After 24 hours mitochondrial membrane depolarization (MΨP) was examined by JC-1 staining by flow cytometry. The data

Techniques Used: Staining, Flow Cytometry, Cytometry

STAT3 is necessary for IL-6-mediated drug resistance. A, CHLA-255 cells were treated with IL-6 (10 ng/mL) and sIL-6R (25 ng/mL) for 30 minutes in the presence of stattic at indicated concentrations. Cell lysates were examined for pSTAT3 and STAT3 expression
Figure Legend Snippet: STAT3 is necessary for IL-6-mediated drug resistance. A, CHLA-255 cells were treated with IL-6 (10 ng/mL) and sIL-6R (25 ng/mL) for 30 minutes in the presence of stattic at indicated concentrations. Cell lysates were examined for pSTAT3 and STAT3 expression

Techniques Used: Expressing

sIL-6R produced by monocytes sensitizes neuroblastoma cells to IL-6-mediated STAT3 activation. A, CHLA-255 cells were treated with IL-6 at indicated concentrations in the absence (top panel) or presence (lower panel) of sIL-6R (25ng/mL) for 30 minutes.
Figure Legend Snippet: sIL-6R produced by monocytes sensitizes neuroblastoma cells to IL-6-mediated STAT3 activation. A, CHLA-255 cells were treated with IL-6 at indicated concentrations in the absence (top panel) or presence (lower panel) of sIL-6R (25ng/mL) for 30 minutes.

Techniques Used: Produced, Activation Assay

IL-6 and sIL-6R activate STAT3 in human neuroblastoma cells. A., The presence of phosphorylated pY 705 STAT3 and STAT3 was detected by Western blot analysis in total cell lysates from 8 neuroblastoma cell lines. When indicated, cells were treated with
Figure Legend Snippet: IL-6 and sIL-6R activate STAT3 in human neuroblastoma cells. A., The presence of phosphorylated pY 705 STAT3 and STAT3 was detected by Western blot analysis in total cell lysates from 8 neuroblastoma cell lines. When indicated, cells were treated with

Techniques Used: Western Blot

30) Product Images from "An inhibitor of interleukin-6 trans-signalling, sgp130, contributes to impaired acute phase response in human chronic liver disease"

Article Title: An inhibitor of interleukin-6 trans-signalling, sgp130, contributes to impaired acute phase response in human chronic liver disease

Journal: Clinical and Experimental Immunology

doi: 10.1111/j.1365-2249.2009.03916.x

Acute phase response mediated by interleukin (IL)-6, assessed by fibrinogen (FBG) measurement in supernatants of 24-h cell cultures: (a) HepG2 cells stimulated with increasing doses of recombinant human IL-6 with or without soluble IL-6 receptor (sIL-6R). HepG2 cells (b) or HuH-7cells (c) stimulated with 50 ng/ml recombinant human IL-6, and increasing doses of gp130–Fc fusion protein, with 100 ng/ml sIL-6R (grey bars) or without sIL-6R (black bars). Values are means ± standard error of the mean of three to five experiments. § P = 0·05; * P
Figure Legend Snippet: Acute phase response mediated by interleukin (IL)-6, assessed by fibrinogen (FBG) measurement in supernatants of 24-h cell cultures: (a) HepG2 cells stimulated with increasing doses of recombinant human IL-6 with or without soluble IL-6 receptor (sIL-6R). HepG2 cells (b) or HuH-7cells (c) stimulated with 50 ng/ml recombinant human IL-6, and increasing doses of gp130–Fc fusion protein, with 100 ng/ml sIL-6R (grey bars) or without sIL-6R (black bars). Values are means ± standard error of the mean of three to five experiments. § P = 0·05; * P

Techniques Used: Recombinant

(a) Circulating interleukin (IL)-6, soluble IL-6 receptor (sIL-6R) and sgp130 levels in 15 healthy subjects (HS) and in 45 patients with alcoholic liver disease, comprising 15 with alcoholic steatohepatitis (ASH) including six with F0–F1 Kleiner fibrosis score and nine F2–F3 fibrosis stage, 15 with alcoholic cirrhosis and 15 with severe alcoholic hepatitis (AH). (b) In 84 HCV infected patients: Metavir fibrosis score F0: n = 9, F1: n = 17, F2: n = 32, F3: n = 10, F4: n = 16; * P
Figure Legend Snippet: (a) Circulating interleukin (IL)-6, soluble IL-6 receptor (sIL-6R) and sgp130 levels in 15 healthy subjects (HS) and in 45 patients with alcoholic liver disease, comprising 15 with alcoholic steatohepatitis (ASH) including six with F0–F1 Kleiner fibrosis score and nine F2–F3 fibrosis stage, 15 with alcoholic cirrhosis and 15 with severe alcoholic hepatitis (AH). (b) In 84 HCV infected patients: Metavir fibrosis score F0: n = 9, F1: n = 17, F2: n = 32, F3: n = 10, F4: n = 16; * P

Techniques Used: Infection

31) Product Images from "Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior"

Article Title: Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior

Journal: Journal of Neuroinflammation

doi: 10.1186/1742-2094-8-54

IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P
Figure Legend Snippet: IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P

Techniques Used:

32) Product Images from "Are Pentraxin 3 and Transsignaling Early Markers for Immunologic Injury Severity in Polytrauma? A Pilot Study"

Article Title: Are Pentraxin 3 and Transsignaling Early Markers for Immunologic Injury Severity in Polytrauma? A Pilot Study

Journal: Clinical Orthopaedics and Related Research

doi: 10.1007/s11999-013-2922-x

The temporal profile of the sIL-6R blood concentration in patients with polytrauma is shown. Low sIL-6R concentrations are seen, without a clear maximum. Horizontal line = time point after admission; box = concentration with SD; whiskers = CI.
Figure Legend Snippet: The temporal profile of the sIL-6R blood concentration in patients with polytrauma is shown. Low sIL-6R concentrations are seen, without a clear maximum. Horizontal line = time point after admission; box = concentration with SD; whiskers = CI.

Techniques Used: Concentration Assay

33) Product Images from "An inhibitor of interleukin-6 trans-signalling, sgp130, contributes to impaired acute phase response in human chronic liver disease"

Article Title: An inhibitor of interleukin-6 trans-signalling, sgp130, contributes to impaired acute phase response in human chronic liver disease

Journal: Clinical and Experimental Immunology

doi: 10.1111/j.1365-2249.2009.03916.x

Acute phase response mediated by interleukin (IL)-6, assessed by fibrinogen (FBG) measurement in supernatants of 24-h cell cultures: (a) HepG2 cells stimulated with increasing doses of recombinant human IL-6 with or without soluble IL-6 receptor (sIL-6R). HepG2 cells (b) or HuH-7cells (c) stimulated with 50 ng/ml recombinant human IL-6, and increasing doses of gp130–Fc fusion protein, with 100 ng/ml sIL-6R (grey bars) or without sIL-6R (black bars). Values are means ± standard error of the mean of three to five experiments. § P = 0·05; * P
Figure Legend Snippet: Acute phase response mediated by interleukin (IL)-6, assessed by fibrinogen (FBG) measurement in supernatants of 24-h cell cultures: (a) HepG2 cells stimulated with increasing doses of recombinant human IL-6 with or without soluble IL-6 receptor (sIL-6R). HepG2 cells (b) or HuH-7cells (c) stimulated with 50 ng/ml recombinant human IL-6, and increasing doses of gp130–Fc fusion protein, with 100 ng/ml sIL-6R (grey bars) or without sIL-6R (black bars). Values are means ± standard error of the mean of three to five experiments. § P = 0·05; * P

Techniques Used: Recombinant

(a) Circulating interleukin (IL)-6, soluble IL-6 receptor (sIL-6R) and sgp130 levels in 15 healthy subjects (HS) and in 45 patients with alcoholic liver disease, comprising 15 with alcoholic steatohepatitis (ASH) including six with F0–F1 Kleiner fibrosis score and nine F2–F3 fibrosis stage, 15 with alcoholic cirrhosis and 15 with severe alcoholic hepatitis (AH). (b) In 84 HCV infected patients: Metavir fibrosis score F0: n = 9, F1: n = 17, F2: n = 32, F3: n = 10, F4: n = 16; * P
Figure Legend Snippet: (a) Circulating interleukin (IL)-6, soluble IL-6 receptor (sIL-6R) and sgp130 levels in 15 healthy subjects (HS) and in 45 patients with alcoholic liver disease, comprising 15 with alcoholic steatohepatitis (ASH) including six with F0–F1 Kleiner fibrosis score and nine F2–F3 fibrosis stage, 15 with alcoholic cirrhosis and 15 with severe alcoholic hepatitis (AH). (b) In 84 HCV infected patients: Metavir fibrosis score F0: n = 9, F1: n = 17, F2: n = 32, F3: n = 10, F4: n = 16; * P

Techniques Used: Infection

34) Product Images from "Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib"

Article Title: Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2019.00541

Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p
Figure Legend Snippet: Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p

Techniques Used: Inhibition

35) Product Images from "IL-6 Trans–Signaling Links Inflammation with Angiogenesis in the Peritoneal Membrane"

Article Title: IL-6 Trans–Signaling Links Inflammation with Angiogenesis in the Peritoneal Membrane

Journal: Journal of the American Society of Nephrology : JASN

doi: 10.1681/ASN.2015101169

IL-6 signaling contributes to VEGF release by HPMCs during peritonitis. (A) IL-6 and sIL-6R levels in dialysates drained from either stable patients on PD after a routine 4-hour dwell ( n =20) or patients with peritonitis at first presentation ( n =11); the data are presented as box and whiskers plots, with the median, 25th and 75th percentiles, and range of data indicated. * P
Figure Legend Snippet: IL-6 signaling contributes to VEGF release by HPMCs during peritonitis. (A) IL-6 and sIL-6R levels in dialysates drained from either stable patients on PD after a routine 4-hour dwell ( n =20) or patients with peritonitis at first presentation ( n =11); the data are presented as box and whiskers plots, with the median, 25th and 75th percentiles, and range of data indicated. * P

Techniques Used:

Identification of sequences in the human VEGF promoter responsive to stimulation with the IL-6 + sIL-6R complex. Cells were transiently transfected with VEGF promoter constructs and stimulated with IL-6 and/or sIL-6R (both at 100 ng/ml) as indicated. Luciferase activity was determined as described in Concise Methods and expressed as relative light units (RLU). (A) Time effect of the IL-6 + sIL-6R complex on the full-length VEGF promoter activity. * P
Figure Legend Snippet: Identification of sequences in the human VEGF promoter responsive to stimulation with the IL-6 + sIL-6R complex. Cells were transiently transfected with VEGF promoter constructs and stimulated with IL-6 and/or sIL-6R (both at 100 ng/ml) as indicated. Luciferase activity was determined as described in Concise Methods and expressed as relative light units (RLU). (A) Time effect of the IL-6 + sIL-6R complex on the full-length VEGF promoter activity. * P

Techniques Used: Transfection, Construct, Luciferase, Activity Assay

Effect of SP4 on VEGF–mediated endothelial cell tube formation. (A) Cells were transiently transfected with either SP4 siRNA or scrambled (scramb.) siRNA, stimulated with IL-6 + sIL-6R (both at 100 ng/ml) for 24 hours, and then, assessed for VEGF secretion. * P
Figure Legend Snippet: Effect of SP4 on VEGF–mediated endothelial cell tube formation. (A) Cells were transiently transfected with either SP4 siRNA or scrambled (scramb.) siRNA, stimulated with IL-6 + sIL-6R (both at 100 ng/ml) for 24 hours, and then, assessed for VEGF secretion. * P

Techniques Used: Transfection

Identification of SP4 as a transcription factor mediating human VEGF promoter induction by the IL-6 + sIL-6R complex. (A and B) Cells were stimulated with IL-6 + sIL-6R at 100 ng/ml for 6 hours, and the nuclear fractions were obtained and analyzed with EMSA. In A, EMSA was performed with consensus oligonucleotide probes for SP1 and SP4. In B, formation of nuclear complexes with SP4 probe was assessed in the presence of either 100-fold molar excess of unlabeled VEGF DNA (resulting in a reduced shift) or SP4-specific antibody (resulting in supershift). (C) Effect of site-directed mutagenesis in the SP4 binding site within the VEGF promoter on its activity after stimulation with IL-6 + sIL-6R. Luciferase activity was expressed as relative light units (RLU). * P
Figure Legend Snippet: Identification of SP4 as a transcription factor mediating human VEGF promoter induction by the IL-6 + sIL-6R complex. (A and B) Cells were stimulated with IL-6 + sIL-6R at 100 ng/ml for 6 hours, and the nuclear fractions were obtained and analyzed with EMSA. In A, EMSA was performed with consensus oligonucleotide probes for SP1 and SP4. In B, formation of nuclear complexes with SP4 probe was assessed in the presence of either 100-fold molar excess of unlabeled VEGF DNA (resulting in a reduced shift) or SP4-specific antibody (resulting in supershift). (C) Effect of site-directed mutagenesis in the SP4 binding site within the VEGF promoter on its activity after stimulation with IL-6 + sIL-6R. Luciferase activity was expressed as relative light units (RLU). * P

Techniques Used: Mutagenesis, Binding Assay, Activity Assay, Luciferase

The role of STAT3 in SP4–mediated VEGF induction by the IL-6 + sIL-6R complex. (A and B) Kinetics of STAT3 and SP4 mRNA induction in HPMCs treated with IL-6 + sIL-6R (both at 100 ng/ml). * P
Figure Legend Snippet: The role of STAT3 in SP4–mediated VEGF induction by the IL-6 + sIL-6R complex. (A and B) Kinetics of STAT3 and SP4 mRNA induction in HPMCs treated with IL-6 + sIL-6R (both at 100 ng/ml). * P

Techniques Used:

Induction of VEGF in HPMCs by a combination of IL-6 and sIL-6R. (A) Kinetics of VEGF secretion by HPMCs treated with IL-6 (10 ng/ml) and sIL-6R (25 ng/ml) either singly or in combination. * P
Figure Legend Snippet: Induction of VEGF in HPMCs by a combination of IL-6 and sIL-6R. (A) Kinetics of VEGF secretion by HPMCs treated with IL-6 (10 ng/ml) and sIL-6R (25 ng/ml) either singly or in combination. * P

Techniques Used:

36) Product Images from "INTERLEUKIN-6 TRANS-SIGNALING SYSTEM IN INTRA-AMNIOTIC INFLAMMATION, PRETERM BIRTH AND PRETERM PREMATURE RUPTURE OF THE MEMBRANES"

Article Title: INTERLEUKIN-6 TRANS-SIGNALING SYSTEM IN INTRA-AMNIOTIC INFLAMMATION, PRETERM BIRTH AND PRETERM PREMATURE RUPTURE OF THE MEMBRANES

Journal: Journal of immunology (Baltimore, Md. : 1950)

doi: 10.4049/jimmunol.1003587

Levels of amniotic fluid (AF) IL-6, sIL-6R, sgp130 and sIL-6R/sgp130 molar ratio in pregnancies complicated by intra-amniotic inflammation (IAI) and preterm premature rupture of the membranes (PPROM)
Figure Legend Snippet: Levels of amniotic fluid (AF) IL-6, sIL-6R, sgp130 and sIL-6R/sgp130 molar ratio in pregnancies complicated by intra-amniotic inflammation (IAI) and preterm premature rupture of the membranes (PPROM)

Techniques Used:

Relationships between amniotic fluid (AF) levels of IL-6, sIL-6R, sgp130, sIL-R/ sgp130 molar ratio and gestational age (GA) in pregnancies with normal outcomes (n=110)
Figure Legend Snippet: Relationships between amniotic fluid (AF) levels of IL-6, sIL-6R, sgp130, sIL-R/ sgp130 molar ratio and gestational age (GA) in pregnancies with normal outcomes (n=110)

Techniques Used:

Gestational regulation of AF trans-signaling molecules IL-6, sIL-6R and sgp130
Figure Legend Snippet: Gestational regulation of AF trans-signaling molecules IL-6, sIL-6R and sgp130

Techniques Used:

Ex-vivo production of MMP-9 and IL-6 following incubation of the fetal membranes with sgp130, sIL-6R and LPS
Figure Legend Snippet: Ex-vivo production of MMP-9 and IL-6 following incubation of the fetal membranes with sgp130, sIL-6R and LPS

Techniques Used: Ex Vivo, Incubation

37) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

38) Product Images from "Circulating Levels of IL‐6 Receptor and gp130 and Long‐Term Clinical Outcomes in ST‐Elevation Myocardial Infarction"

Article Title: Circulating Levels of IL‐6 Receptor and gp130 and Long‐Term Clinical Outcomes in ST‐Elevation Myocardial Infarction

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

doi: 10.1161/JAHA.115.003014

Time‐to‐event curves for the highest quartile (Q4) of soluble interleukin‐6 receptor ( sIL ‐6R) vs the 3 lowest quartiles (Q1–Q3) measured in 989  STEMI  patients according to the primary endpoint. The log rank test was used to compare the survival curves. STEMI indicates ST‐elevation myocardial infarction.
Figure Legend Snippet: Time‐to‐event curves for the highest quartile (Q4) of soluble interleukin‐6 receptor ( sIL ‐6R) vs the 3 lowest quartiles (Q1–Q3) measured in 989 STEMI patients according to the primary endpoint. The log rank test was used to compare the survival curves. STEMI indicates ST‐elevation myocardial infarction.

Techniques Used:

Time‐to‐event curves for the highest quartile (Q4) of soluble interleukin‐6 receptor ( sIL ‐6R) vs the 3 lowest quartiles (Q1–Q3) measured in 989  STEMI  patients according to the secondary endpoint. The log rank test was used to compare the survival curves. STEMI indicates ST‐elevation myocardial infarction.
Figure Legend Snippet: Time‐to‐event curves for the highest quartile (Q4) of soluble interleukin‐6 receptor ( sIL ‐6R) vs the 3 lowest quartiles (Q1–Q3) measured in 989 STEMI patients according to the secondary endpoint. The log rank test was used to compare the survival curves. STEMI indicates ST‐elevation myocardial infarction.

Techniques Used:

39) Product Images from "Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib"

Article Title: Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2019.00541

Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p
Figure Legend Snippet: Effect of tofacitinib and baricitinib on IL-6 dependent release of cytokines. (A) RASF were pretreated with JAKi or vehicle control (DMSO 0.1%) for 2 h and then additionally activated with OSM (100 ng/ml) for 24 h. The IL-6 release was decreased by tofacitinib or baricitinib confirming the inhibition of JAK dependent pathway. (B) The addition of sIL-6R to IL-1β stimulated RASF leads to an increase of IL-6 but not of IL-8 release. This increase could be blocked by tofacitinib. * p

Techniques Used: Inhibition

40) Product Images from "Microglia priming by interleukin-6 signaling is enhanced in aged mice"

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

Journal: Journal of neuroimmunology

doi: 10.1016/j.jneuroim.2018.09.002

Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p
Figure Legend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

Techniques Used: Enzyme-linked Immunosorbent Assay

Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p
Figure Legend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

Techniques Used: Expressing

sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p
Figure Legend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

Techniques Used: Expressing, Dot Blot, Fluorescence

Related Articles

other:

Article Title: Inhibition of interleukin-6 trans-signaling prevents inflammation and endothelial barrier disruption in retinal endothelial cells.
Article Snippet: Pre-treatment with sgp130Fc before IL-6/sIL-6R exposure significantly reduced the phosphorylation of STAT3 in comparison to the IL-6/sIL-6R treated group.

Article Title: Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior
Article Snippet: Here, STAT3 was upregulated in response to both IL-6 and LPS in BV.2 and Neuro.2A cells and pretreatment with sIL-6R led to an increased IL-6- and LPS-induced STAT3 phosphorylation.

Recombinant:

Article Title: Interleukin-6/interleukin-6 receptor complex promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells
Article Snippet: Exogenous stimulation assay AG490, a specific STAT3 signaling pathway inhibitor, was added at a concentration of 10 μM during the osteogenic differentiation of MSCs. .. Recombinant human IL-6 protein (100 ng/ml), recombinant human sIL-6R protein (100 ng/ml), anti-IL-6 antibody (5 μg/ml) or anti-IL-6R antibody (5 μg/ml) was added to determine the roles of IL-6/IL-6R in the osteogenic differentiation of MSCs (all from R & D Systems). .. Lentiviruses construction and infection Lentiviruses encoding short hairpin RNA (shRNA) for IL-6 were constructed with a target sequence of 5′- GCAGGACATGACAACTCATCT-3′ (Lv-IL6), and shRNA for IL-6R were also constructed with a target sequence of 5′- GGAAGACAATGCCACTGTTCA-3′ (Lv-IL6R).

Article Title: IL-27 mediates HLA class I up-regulation, which can be inhibited by the IL-6 pathway, in HLA-deficient Small Cell Lung Cancer cells
Article Snippet: .. For immunofluorescence and QRT-PCR analyses, cells were seeded in 24-well plates in culture medium at 5 × 104 cells/well and different cytokines were added: IFN-γ (1000 IU/ml, PeproTech, 300–02), IL-27 (100 ng/ml R & D System, 2526-IL-010), IL-6 (50 ng/ml R & D System 206-IL-010) or recombinant human IL-6Rα/IL-6 chimera [sIL-6R/IL-6] (50 ng/ml R & D System 8954-SR-025). .. Treatments were carried out for 48 h. For the analysis of tyrosine-phosphorylated STAT proteins, 1 × 105 SCLC cells were incubated in a test tube at 37 °C with or without 50 ng/ ml of IL-27, 20 ng/ml of IL-6, 40 ng/ml of sIL-6R/IL-6 in 0.5 ml of medium for the 10, 30 or 60 min time points.

Mouse Assay:

Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice
Article Snippet: We therefore examined differences in signaling molecules between adult and aged mice in the absence of inflammatory stimulus. .. The cerebrospinal fluid of aged mice had increased amounts of sIL-6R compared with adult counterparts, although adult and aged mice did not produce significantly different amounts of its inhibitor, sgp130 ( ). .. Furthermore, the brains of aged mice had elevated amounts of Cd11b+ /MHC-II+ cells, indicative of primed microglia.

Enzyme-linked Immunosorbent Assay:

Article Title: Maternal country of origin, breast milk characteristics and potential influences on immunity in offspring
Article Snippet: .. sCD14, CXCL-8/IL-8, TGF-β1, TGF-β2, sIL-6R, sgp130 and total IgA levels in breast milk and in culture supernatants from IECs (HT-29) were determined using ELISA (sCD14, CXCL-8/IL-8, TGF-β1, TGF-β2, sIL-6R and sgp130 ELISAs from R & D Systems, Abingdon, Oxon, UK and human IgA ELISA quantitation kit from Bethyl Laboratories Inc., Montgomery, TX, USA), according to the manufacturer's instructions. .. The optical density was determined using a microplate reader (Molecular Devices, Sunnyvale, CA, USA) set at 450 nm.

Quantitation Assay:

Article Title: Maternal country of origin, breast milk characteristics and potential influences on immunity in offspring
Article Snippet: .. sCD14, CXCL-8/IL-8, TGF-β1, TGF-β2, sIL-6R, sgp130 and total IgA levels in breast milk and in culture supernatants from IECs (HT-29) were determined using ELISA (sCD14, CXCL-8/IL-8, TGF-β1, TGF-β2, sIL-6R and sgp130 ELISAs from R & D Systems, Abingdon, Oxon, UK and human IgA ELISA quantitation kit from Bethyl Laboratories Inc., Montgomery, TX, USA), according to the manufacturer's instructions. .. The optical density was determined using a microplate reader (Molecular Devices, Sunnyvale, CA, USA) set at 450 nm.

Blocking Assay:

Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation
Article Snippet: .. Most importantly, blocking sIL-6R with sIL-6R-specific neutralizing antibody completely abolished this growth-promoting effect ( P < 0.01, ) but a matched isotype control antibody could not block the effect. ..

Immunofluorescence:

Article Title: IL-27 mediates HLA class I up-regulation, which can be inhibited by the IL-6 pathway, in HLA-deficient Small Cell Lung Cancer cells
Article Snippet: .. For immunofluorescence and QRT-PCR analyses, cells were seeded in 24-well plates in culture medium at 5 × 104 cells/well and different cytokines were added: IFN-γ (1000 IU/ml, PeproTech, 300–02), IL-27 (100 ng/ml R & D System, 2526-IL-010), IL-6 (50 ng/ml R & D System 206-IL-010) or recombinant human IL-6Rα/IL-6 chimera [sIL-6R/IL-6] (50 ng/ml R & D System 8954-SR-025). .. Treatments were carried out for 48 h. For the analysis of tyrosine-phosphorylated STAT proteins, 1 × 105 SCLC cells were incubated in a test tube at 37 °C with or without 50 ng/ ml of IL-27, 20 ng/ml of IL-6, 40 ng/ml of sIL-6R/IL-6 in 0.5 ml of medium for the 10, 30 or 60 min time points.

Quantitative RT-PCR:

Article Title: IL-27 mediates HLA class I up-regulation, which can be inhibited by the IL-6 pathway, in HLA-deficient Small Cell Lung Cancer cells
Article Snippet: .. For immunofluorescence and QRT-PCR analyses, cells were seeded in 24-well plates in culture medium at 5 × 104 cells/well and different cytokines were added: IFN-γ (1000 IU/ml, PeproTech, 300–02), IL-27 (100 ng/ml R & D System, 2526-IL-010), IL-6 (50 ng/ml R & D System 206-IL-010) or recombinant human IL-6Rα/IL-6 chimera [sIL-6R/IL-6] (50 ng/ml R & D System 8954-SR-025). .. Treatments were carried out for 48 h. For the analysis of tyrosine-phosphorylated STAT proteins, 1 × 105 SCLC cells were incubated in a test tube at 37 °C with or without 50 ng/ ml of IL-27, 20 ng/ml of IL-6, 40 ng/ml of sIL-6R/IL-6 in 0.5 ml of medium for the 10, 30 or 60 min time points.

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    R&D Systems sil 6r
    Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor <t>(sIL-6R)</t> expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
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    Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

    Journal: Carcinogenesis

    Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

    doi: 10.1093/carcin/bgr075

    Figure Lengend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

    Article Snippet: Most importantly, blocking sIL-6R with sIL-6R-specific neutralizing antibody completely abolished this growth-promoting effect ( P < 0.01, ) but a matched isotype control antibody could not block the effect.

    Techniques: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

    Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

    Journal: Carcinogenesis

    Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

    doi: 10.1093/carcin/bgr075

    Figure Lengend Snippet: Blocking the IL-6/sIL-6R trans-signaling axis affects the growth/survival of MSLN expressing PC cell proliferation. ( A ) sIL-6R antibody blocking can reduce the endogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody. The cells were then treated with 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability after a treatment by viability of same cells without treatment (or dimethyl sulfoxide treated) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. (B). Soluble IL-6R antibody blocking can reduce the exogenous IL-6-induced proliferation of MIA-MSLN cells. MIA-MSLN cells were serum-starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. ( C ) IL6/sIL-6R trans-signaling axis is operative in MIA cell proliferation. Both MIA-V and MIA-MSLN cells were serum starved (0% FBS) for 24 h with or without anti-sIL-6R or isotype control antibody and then treated with a cocktail of rIL-6/rsIL-6R in 0.2% FBS containing media for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability of treated cells (treated with rIL-6/rsIL-6R cocktail with or without anti-sIL-6R antibody blocking) by that of untreated cells and is plotted along Y -axis. Data plotted show mean of triplicate wells. *, # denote P

    Article Snippet: Most importantly, blocking sIL-6R with sIL-6R-specific neutralizing antibody completely abolished this growth-promoting effect ( P < 0.01, ) but a matched isotype control antibody could not block the effect.

    Techniques: Blocking Assay, Expressing, MTT Assay

    Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

    Journal: Journal of neuroimmunology

    Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

    doi: 10.1016/j.jneuroim.2018.09.002

    Figure Lengend Snippet: Pro-inflammatory soluble IL-6 receptor, but not soluble gp130, is increased in aged animals. ELISA was performed on Cerebral Spinal Fluid (CSF) collected from the cisterna magna of adult and aged animals (n=6–7; samples plated in duplicate). The CSF of naïve aged animals contained increased amounts of sIL6-R compared to their adult counterparts. Amounts of sgp130 were not different between groups. Means with different letters are significantly different from each other (p

    Article Snippet: The cerebrospinal fluid of aged mice had increased amounts of sIL-6R compared with adult counterparts, although adult and aged mice did not produce significantly different amounts of its inhibitor, sgp130 ( ).

    Techniques: Enzyme-linked Immunosorbent Assay

    Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

    Journal: Journal of neuroimmunology

    Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

    doi: 10.1016/j.jneuroim.2018.09.002

    Figure Lengend Snippet: Effects of sIL-6R on cytokine gene expression in BV.2 microglia. BV.2 cells (n=3–5 replicates) were treated with (A-C) IL-6 (100pg/mL) or (D-F) LPS (100ng/mL) for 8hr in the presence or absence of sIL-6R (25ng/mL, 1hr pre-treated). Cytokine gene expression was significantly increased in the presence of sIL-6R after treatment with both IL-6 and LPS in all genes. sIL-6R was required for changes in gene expression of (A) IL-1β and (B) I L-10 after treatment with IL-6. Although LPS was able to induce changes in gene expression in the absence of sIL-6R ( D-F), the presence of sIL-6R greatly enhanced these changes. Means with different letters are statistically different from each other (p

    Article Snippet: The cerebrospinal fluid of aged mice had increased amounts of sIL-6R compared with adult counterparts, although adult and aged mice did not produce significantly different amounts of its inhibitor, sgp130 ( ).

    Techniques: Expressing

    sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

    Journal: Journal of neuroimmunology

    Article Title: Microglia priming by interleukin-6 signaling is enhanced in aged mice

    doi: 10.1016/j.jneuroim.2018.09.002

    Figure Lengend Snippet: sIL-6R enhances IL-6 induced MHC-II expression in BV.2 microglia. (A) Representative dot blot of MHCI-II expression from BV.2 microglial cells (n=3–5 replicates) treated with IL-6 (12hr) in the presence or absence of sIL-6R (25ng/mL; 1 hr pre-) (bottom panel). (B) MHC-II expression is enhanced when cells are treated with 100pg/mL of IL-6 in the presence of sIL-6R, but not 100pg/mL of IL-6 alone. A ceiling effect is reached at 1000 pg/mL of IL-6, regardless of the presence of sIL-6R. (C) This ceiling effect is confirmed by mean fluorescence intensity (MFI) data. Means with different letters are significantly different from each other (p

    Article Snippet: The cerebrospinal fluid of aged mice had increased amounts of sIL-6R compared with adult counterparts, although adult and aged mice did not produce significantly different amounts of its inhibitor, sgp130 ( ).

    Techniques: Expressing, Dot Blot, Fluorescence

    IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P

    Journal: Journal of Neuroinflammation

    Article Title: Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior

    doi: 10.1186/1742-2094-8-54

    Figure Lengend Snippet: IL-6 trans-signaling in BV.2 microglia and Neuro.2A cells . BV.2 and Neuro2A cells were pre-treated for 1 h with 25 ng/mL sIL-6R and A) IL-6-induced STAT3 phosphorylation and B) LPS-induced IL-6 protein secretion were measured at 20 min and 3 h, respectively. Results are an average of 5 independent experiments. Means with different letters are significantly different from one another (P

    Article Snippet: Here, STAT3 was upregulated in response to both IL-6 and LPS in BV.2 and Neuro.2A cells and pretreatment with sIL-6R led to an increased IL-6- and LPS-induced STAT3 phosphorylation.

    Techniques: