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InvivoGen pam2csk 4
Pam2csk 4, supplied by InvivoGen, used in various techniques. Bioz Stars score: 96/100, based on 605 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pam2csk+4/pmc12768884-35-1-8?v=InvivoGen
Average 96 stars, based on 605 article reviews

pam2csk 4 - by Bioz Stars, 2026-08

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a Timeline of TLR2 tolerance induction. Timeline for the 5-week induction of demyelination with 0.2% cuprizone (or normal chow), followed by the 2-week remyelination period on normal chow. WT mice were treated with either low dose P2C or VC during the remyelination period. Mice were sacrificed and analyzed after the 2-week remyelination period. b Timeline of TLR2 −/− . Timeline for the 5-week induction of demyelination with 0.2% cuprizone (or normal chow) followed by the 2-week remyelination period. WT and TLR2 −/− mice were not treated during the remyelination period. Mice were sacrificed and analyzed immediately following the 5-week cuprizone (or normal chow) feeding to evaluate baseline demyelination levels, or after the 2-week remyelination period on normal chow to evaluate myelin recovery. c Systemic TLR2 tolerance induction. At the end of the 2-week remyelination period, WT mice that had been fed for the first 5 weeks with either cuprizone or normal chow and treated with P2C or VC as in a received a single i.v. injection of Pam3CSK 4 (P3C, 100 μg) and 2 h later serum was obtained and analyzed for TNFα by ELISA. Results are expressed as mean values per mouse. N = 9 mice/cohort. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA. **** p < 0.0001. For NS comparisons, VC treated: normal vs cuprizone-fed, p = 0.8118. P2C treated: normal vs cuprizone-fed, p = 0.9972

Journal: Journal of Neuroinflammation

Article Title: Systemic TLR2 tolerance enhances central nervous system remyelination

doi: 10.1186/s12974-019-1540-2

Figure Lengend Snippet: a Timeline of TLR2 tolerance induction. Timeline for the 5-week induction of demyelination with 0.2% cuprizone (or normal chow), followed by the 2-week remyelination period on normal chow. WT mice were treated with either low dose P2C or VC during the remyelination period. Mice were sacrificed and analyzed after the 2-week remyelination period. b Timeline of TLR2 −/− . Timeline for the 5-week induction of demyelination with 0.2% cuprizone (or normal chow) followed by the 2-week remyelination period. WT and TLR2 −/− mice were not treated during the remyelination period. Mice were sacrificed and analyzed immediately following the 5-week cuprizone (or normal chow) feeding to evaluate baseline demyelination levels, or after the 2-week remyelination period on normal chow to evaluate myelin recovery. c Systemic TLR2 tolerance induction. At the end of the 2-week remyelination period, WT mice that had been fed for the first 5 weeks with either cuprizone or normal chow and treated with P2C or VC as in a received a single i.v. injection of Pam3CSK 4 (P3C, 100 μg) and 2 h later serum was obtained and analyzed for TNFα by ELISA. Results are expressed as mean values per mouse. N = 9 mice/cohort. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA. **** p < 0.0001. For NS comparisons, VC treated: normal vs cuprizone-fed, p = 0.8118. P2C treated: normal vs cuprizone-fed, p = 0.9972

Article Snippet: Starting on day 33, mice received intravenous (i.v.) injections of PBS vehicle control (VC) or Pam2CSK 4 (P2C) (2.5 μg) (InvivoGen; San Diego, CA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Effect of TLR2 tolerance induction on OPC and OL frequency. a – d Overlapping immunohistofluorescent stains of one representative corpus callosum section. a Green = PDGFRα + cells, b red = CC1 + cells, c magenta = Olig2 + cells, d all images merged; blue = DAPI. In d , the white arrow depicts the example CC1 + cell also seen in b , and the green arrow depicts the example PDGFRα + cell also seen in a . Scale bars represent 50 μm. e–g Analysis of cell frequency in corpus callosum sections based on co-localization of DAPI + Olig2 + cells/mm 2 . e Olig2 + cells/mm 2 , f Olig2 + CC1 + cells/mm 2 , g Olig2 + PDGFRα + cells/mm 2 . Data points represent individually analyzed sections derived from 4 mice per cohort; 3–4 sections were analyzed from each mouse. N = 12–15 total sections per experimental cohort. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA. * p < 0.05; ** p < 0.01. For NS comparisons: e VC treated: normal vs cuprizone-fed, p = 0.1660. P2C treated: normal vs cuprizone-fed, p = 0.7672. Normal chow: VC vs P2C, p = 0.5803. Cuprizone-fed: VC vs P2C, p = 0.9451. f VC treated: normal vs cuprizone-fed, p = 0.1295. P2C treated: normal vs cuprizone-fed, p = 0.7142. Normal chow: VC vs P2C, p = 0.6692. Cuprizone-fed: VC vs P2C, p = 0.9772. g Cuprizone-fed: VC vs P2C, p = 0.9983

Journal: Journal of Neuroinflammation

Article Title: Systemic TLR2 tolerance enhances central nervous system remyelination

doi: 10.1186/s12974-019-1540-2

Figure Lengend Snippet: Effect of TLR2 tolerance induction on OPC and OL frequency. a – d Overlapping immunohistofluorescent stains of one representative corpus callosum section. a Green = PDGFRα + cells, b red = CC1 + cells, c magenta = Olig2 + cells, d all images merged; blue = DAPI. In d , the white arrow depicts the example CC1 + cell also seen in b , and the green arrow depicts the example PDGFRα + cell also seen in a . Scale bars represent 50 μm. e–g Analysis of cell frequency in corpus callosum sections based on co-localization of DAPI + Olig2 + cells/mm 2 . e Olig2 + cells/mm 2 , f Olig2 + CC1 + cells/mm 2 , g Olig2 + PDGFRα + cells/mm 2 . Data points represent individually analyzed sections derived from 4 mice per cohort; 3–4 sections were analyzed from each mouse. N = 12–15 total sections per experimental cohort. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA. * p < 0.05; ** p < 0.01. For NS comparisons: e VC treated: normal vs cuprizone-fed, p = 0.1660. P2C treated: normal vs cuprizone-fed, p = 0.7672. Normal chow: VC vs P2C, p = 0.5803. Cuprizone-fed: VC vs P2C, p = 0.9451. f VC treated: normal vs cuprizone-fed, p = 0.1295. P2C treated: normal vs cuprizone-fed, p = 0.7142. Normal chow: VC vs P2C, p = 0.6692. Cuprizone-fed: VC vs P2C, p = 0.9772. g Cuprizone-fed: VC vs P2C, p = 0.9983

Article Snippet: Starting on day 33, mice received intravenous (i.v.) injections of PBS vehicle control (VC) or Pam2CSK 4 (P2C) (2.5 μg) (InvivoGen; San Diego, CA).

Techniques: Derivative Assay

Effect of TLR2 tolerance induction on microglia frequency and phenotype. a – d overlapping immunohistofluorescent stains of one representative corpus callosum section. a green = Arg1 + cells, b red = IBA1 + cells; c magenta = iNOS + cells; d all images merged; blue = DAPI. In d , the green arrow depicts the example Arg1 + cell also shown in a , the yellow arrow depicts the example iNOS + cell also seen in c , and the white arrow depicts the example double-positive, iNOS + Arg1 + cell, also seen in a and c . Scale bars represent 50 μm. e Analysis of IBA1 + cell frequency in corpus callosum sections, based on co-localization of DAPI + IBA1 + cells. f Percentage of IBA1 + cells that are iNOS + (total = single and double positives). g Percentage of IBA1 + cells that are Arg1 + (total = single and double positives). h Percentage of IBA1 + cells that are iNOS + single positive (Arg1 − ). i Percentage of IBA1 + cells that are Arg1 + single positive (iNOS − ). j Percentage of IBA1 + cells that are iNOS + Arg1 + double-positive. Data points represent individually analyzed sections derived from 4 mice per cohort; 2–3 sections were analyzed from each mouse. N = 8–12 total sections per experimental cohort. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. For NS comparisons: e Cuprizone-fed: VC vs P2C, p = 0.9042. f P2C treated: normal vs cuprizone-fed, p = 0.3729. g VC treated: normal vs cuprizone-fed, p = 0.6316. h P2C treated: normal vs cuprizone-fed, p > 0.9999. j VC treated: normal vs cuprizone-fed, p = 0.6881. P2C treated: normal vs cuprizone-fed, p = 0.1123. Cuprizone-fed: VC vs P2C, p = 0.9999

Journal: Journal of Neuroinflammation

Article Title: Systemic TLR2 tolerance enhances central nervous system remyelination

doi: 10.1186/s12974-019-1540-2

Figure Lengend Snippet: Effect of TLR2 tolerance induction on microglia frequency and phenotype. a – d overlapping immunohistofluorescent stains of one representative corpus callosum section. a green = Arg1 + cells, b red = IBA1 + cells; c magenta = iNOS + cells; d all images merged; blue = DAPI. In d , the green arrow depicts the example Arg1 + cell also shown in a , the yellow arrow depicts the example iNOS + cell also seen in c , and the white arrow depicts the example double-positive, iNOS + Arg1 + cell, also seen in a and c . Scale bars represent 50 μm. e Analysis of IBA1 + cell frequency in corpus callosum sections, based on co-localization of DAPI + IBA1 + cells. f Percentage of IBA1 + cells that are iNOS + (total = single and double positives). g Percentage of IBA1 + cells that are Arg1 + (total = single and double positives). h Percentage of IBA1 + cells that are iNOS + single positive (Arg1 − ). i Percentage of IBA1 + cells that are Arg1 + single positive (iNOS − ). j Percentage of IBA1 + cells that are iNOS + Arg1 + double-positive. Data points represent individually analyzed sections derived from 4 mice per cohort; 2–3 sections were analyzed from each mouse. N = 8–12 total sections per experimental cohort. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. For NS comparisons: e Cuprizone-fed: VC vs P2C, p = 0.9042. f P2C treated: normal vs cuprizone-fed, p = 0.3729. g VC treated: normal vs cuprizone-fed, p = 0.6316. h P2C treated: normal vs cuprizone-fed, p > 0.9999. j VC treated: normal vs cuprizone-fed, p = 0.6881. P2C treated: normal vs cuprizone-fed, p = 0.1123. Cuprizone-fed: VC vs P2C, p = 0.9999

Article Snippet: Starting on day 33, mice received intravenous (i.v.) injections of PBS vehicle control (VC) or Pam2CSK 4 (P2C) (2.5 μg) (InvivoGen; San Diego, CA).

Techniques: Derivative Assay

In vitro microglia TLR2 tolerance. Mixed glial cultures were established from P1–P3 WT mouse pups. After 12–18 days in culture, non-adherent cells (microglia) were harvested and either directly plated for testing in 96-well plates or plated after further purification by FACS-sorting for CD45.2 + CD11b + microglia. Microglia were plated at 2–7 × 10 4 cells/well in 96-well plates pre-coated with poly-L lysine. Microglial cultures were designated as either “non-stimulated,” “stimulated,” or “tolerized”. “Non-stimulated” wells received no stimulation over the entire 48 h in culture. “Stimulated” wells received 1 μg/mL P2C only at the 24 h point of culture. “Tolerized” wells received 1 μg/mL P2C both at the initiation of culture (hour 0) and again at the 24 h point of culture. After 48 h, supernatants were harvested and assayed for cytokines via multiplex ELISA. Cytokine levels in “non-stimulated” culture supernatants were either very low or below the level of detection (see Additional file : Table S1). Results are expressed as fold change of cytokine levels in “stimulated” versus “tolerized” culture supernatants for a TNFα; b IL-6; c IL-10. N = 6 total experiments; 2 using non-FACS-sorted microglia (65–75% CD45 + CD11b + cells) and 4 using FACS-sorted microglia. Error bars represent the mean ± SEM. Statistical differences were assessed by Mann-Whitney analysis. * p < 0.05

Journal: Journal of Neuroinflammation

Article Title: Systemic TLR2 tolerance enhances central nervous system remyelination

doi: 10.1186/s12974-019-1540-2

Figure Lengend Snippet: In vitro microglia TLR2 tolerance. Mixed glial cultures were established from P1–P3 WT mouse pups. After 12–18 days in culture, non-adherent cells (microglia) were harvested and either directly plated for testing in 96-well plates or plated after further purification by FACS-sorting for CD45.2 + CD11b + microglia. Microglia were plated at 2–7 × 10 4 cells/well in 96-well plates pre-coated with poly-L lysine. Microglial cultures were designated as either “non-stimulated,” “stimulated,” or “tolerized”. “Non-stimulated” wells received no stimulation over the entire 48 h in culture. “Stimulated” wells received 1 μg/mL P2C only at the 24 h point of culture. “Tolerized” wells received 1 μg/mL P2C both at the initiation of culture (hour 0) and again at the 24 h point of culture. After 48 h, supernatants were harvested and assayed for cytokines via multiplex ELISA. Cytokine levels in “non-stimulated” culture supernatants were either very low or below the level of detection (see Additional file : Table S1). Results are expressed as fold change of cytokine levels in “stimulated” versus “tolerized” culture supernatants for a TNFα; b IL-6; c IL-10. N = 6 total experiments; 2 using non-FACS-sorted microglia (65–75% CD45 + CD11b + cells) and 4 using FACS-sorted microglia. Error bars represent the mean ± SEM. Statistical differences were assessed by Mann-Whitney analysis. * p < 0.05

Article Snippet: Starting on day 33, mice received intravenous (i.v.) injections of PBS vehicle control (VC) or Pam2CSK 4 (P2C) (2.5 μg) (InvivoGen; San Diego, CA).

Techniques: In Vitro, Purification, Multiplex Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Effect of TLR2 tolerance induction on remyelination. a – d Representative EM images from corpus callosum: a “Normal Chow VC”—mice fed for 7 weeks with normal chow and treated for the last 2 weeks with VC. b “Cuprizone VC”—mice fed for 5 weeks with cuprizone, then fed for 2 weeks with normal chow and treated for these last 2 weeks with VC. c “Normal Chow P2C”—mice fed for 7 weeks with normal chow and treated for the last 2 weeks with Pam2CSK 4 (P2C). d “Cuprizone P2C”—mice fed for 5 weeks with cuprizone, then fed for 2 weeks with normal chow and treated for these last 2 weeks with P2C. Scale bar = 2 μm. Red arrows depict typical axons with normal myelin thickness. Yellow arrows depict typical axons with decreased myelin thickness. e Percentage of unmyelinated axons as calculated from EM images. f Myelin thickness (g-ratios) as calculated from EM images. For e and f , data points represent individual mice analyzed as described in the “ ” section. N = 8–10 mice per experimental condition. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA. *** p < 0.001; **** p < 0.0001. For NS comparisons: P2C treated: normal vs cuprizone-fed, p = 0.7912 ( e ). P2C treated: normal vs cuprizone-fed, p = 0.1486 ( f )

Journal: Journal of Neuroinflammation

Article Title: Systemic TLR2 tolerance enhances central nervous system remyelination

doi: 10.1186/s12974-019-1540-2

Figure Lengend Snippet: Effect of TLR2 tolerance induction on remyelination. a – d Representative EM images from corpus callosum: a “Normal Chow VC”—mice fed for 7 weeks with normal chow and treated for the last 2 weeks with VC. b “Cuprizone VC”—mice fed for 5 weeks with cuprizone, then fed for 2 weeks with normal chow and treated for these last 2 weeks with VC. c “Normal Chow P2C”—mice fed for 7 weeks with normal chow and treated for the last 2 weeks with Pam2CSK 4 (P2C). d “Cuprizone P2C”—mice fed for 5 weeks with cuprizone, then fed for 2 weeks with normal chow and treated for these last 2 weeks with P2C. Scale bar = 2 μm. Red arrows depict typical axons with normal myelin thickness. Yellow arrows depict typical axons with decreased myelin thickness. e Percentage of unmyelinated axons as calculated from EM images. f Myelin thickness (g-ratios) as calculated from EM images. For e and f , data points represent individual mice analyzed as described in the “ ” section. N = 8–10 mice per experimental condition. Error bars represent the mean ± SEM. Statistical differences were assessed by 2-way ANOVA. *** p < 0.001; **** p < 0.0001. For NS comparisons: P2C treated: normal vs cuprizone-fed, p = 0.7912 ( e ). P2C treated: normal vs cuprizone-fed, p = 0.1486 ( f )

Article Snippet: Starting on day 33, mice received intravenous (i.v.) injections of PBS vehicle control (VC) or Pam2CSK 4 (P2C) (2.5 μg) (InvivoGen; San Diego, CA).

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