Supplementary Materials

Supplementary Materials for:

Switching of the Relative Dominance Between Feedback Mechanisms in Lipopolysaccharide-Induced NF-κB Signaling

Myong-Hee Sung,* Ning Li, Qizong Lao, Rachel A. Gottschalk, Gordon L. Hager,* Iain D. C. Fraser*

*Corresponding author. E-mail: sungm@mail.nih.gov (M.-H.S.); hagerg@mail.nih.gov (G.L.H.); fraseri@niaid.nih.gov (I.D.C.F.)

This PDF file includes:

  • Fig. S1. Design of reporter constructs and generation of the stable cell clone.
  • Fig. S2. Time series of mCherry intensities and mCherry synthesis rates and analysis of the poor correlation between static measurements of NF-κB and target reporter activities.
  • Fig. S3. Time courses of NF-κB dynamics and reporter activities in single cells.
  • Fig. S4. The time course of the ratio of nuclear NF-κB to total NF-κB deviates from that of the mean nuclear intensity of NF-κB.
  • Fig. S5. LPS dose–sensitive increase of EGFP-RelA in RAW264.7 reporter cells.
  • Fig. S6. Partial least squares regression identifies the nuclear occupancy of RelA as the most predictive variable for mCherry reporter activity in RAW264.7 reporter cells.
  • Fig. S7. LPS dose–dependent induction of Rela in RAW264.7 cells and macrophages.
  • Fig. S8. Gene expression microarray analysis of primary macrophages reveals that a functional transcriptional program is induced only by a high dose of LPS.
  • Fig. S9. Numerous NF-κB positive feedback genes are activated specifically by a high dose of LPS, shifting the balance in the system toward the prevalence of positive feedback.
  • Fig. S10. Ikaros as a candidate factor for the macrophage-specific induction of RelA expression in response to a high dose of LPS.
  • Table S1. Parameter values for mathematical modeling and computation.
  • Table S2. Initial condition for the delay differential equations.
  • Legends for movies S1 to S7

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 4.22 MB

Other Supplementary Material for this manuscript includes the following:

  • File 1. MATLAB code for mathematical modeling.
  • Movie S1 (.mov format). EGFP-RelA kinetics in RAW264.7 cells treated with LPS (10 ng/ml).
  • Movie S2 (.mov format). Kinetics of Tnf promoter–driven mCherry expression in RAW264.7 cells treated with LPS (10 ng/ml).
  • Movie S3 (.mov format). Kinetics of both EGFP-RelA and Tnf promoter–driven mCherry in RAW264.7 cells treated with LPS (10 ng/ml).
  • Movie S4 (.mov format). Kinetics of both EGFP-RelA and Tnf promoter–driven mCherry in RAW264.7 cells treated with LPS (2.5 ng/ml).
  • Movie S5 (.mov format). Kinetics of both EGFP-RelA and Tnf promoter–driven mCherry in RAW264.7 cells treated with LPS (0.5 ng/ml).
  • Movie S6 (.mov format). Kinetics of both EGFP-RelA and Tnf promoter–driven mCherry in RAW264.7 cells treated with LPS (0.1 ng/ml).
  • Movie S7 (.mov format). Kinetics of both EGFP-RelA and Tnf promoter–driven mCherry in untreated RAW264.7 cells.

Citation: M.-H.Sung, N. Li, Q. Lao, R. A. Gottschalk, G. L. Hager, I. D. C. Fraser, Switching of the Relative Dominance Between Feedback Mechanisms in Lipopolysaccharide-Induced NF-κB Signaling. Sci. Signal. 7, ra6 (2014).

© 2014 American Association for the Advancement of Science