Flowchart outlining the steps in the gain-of-function and loss-of-function genomic screens

The gain-of-function screen was used to identify positive modifiers of the NFκB signaling pathway and the loss-of-function screen was used to identify negative modifiers. Approximately 14,500 full-length human and mouse genes were screened for activity.<p><b>Copyright information:</b></p><p>Taken from "A functional map of NFκB signaling identifies novel modulators and multiple system controls"</p><p>http://genomebiology.com/2007/8/6/R104</p><p>Genome Biology 2007;8(6):R104-R104.</p><p>Published online 6 Jun 2007</p><p>PMCID:PMC2394752.</p><p></p

Functional classification of the NFκB modulators identified in the functional genomic screens

Functional classification was performed using NIH David 2.1. Classification of the positive modulators. Classification of the negative modulators.<p><b>Copyright information:</b></p><p>Taken from "A functional map of NFκB signaling identifies novel modulators and multiple system controls"</p><p>http://genomebiology.com/2007/8/6/R104</p><p>Genome Biology 2007;8(6):R104-R104.</p><p>Published online 6 Jun 2007</p><p>PMCID:PMC2394752.</p><p></p

Electrochemical Reduction of Flue Gas Denitrification Wastewater to Ammonia Using a Dual-Defective Cu₂O@Cu Heterojunction Electrode

Wet flue gas denitrification offers a new route to convert industrial nitrogen oxides (NOx) into highly concentrated nitrate wastewater, from which the nitrogen resource can be recovered to ammonia (NH3) via electrochemical nitrate reduction reactions (NITRRs). Low-cost, scalable, and efficient cathodic materials need to be developed to enhance the NH3 production rate. Here, in situ electrodeposition was adopted to fabricate a foamy Cu-based heterojunction electrode containing both Cu-defects and oxygen vacancy loaded Cu2O (OVs-Cu2O), which achieved an NH3 yield rate of 3.59 mmol h–1 cm–2, NH3 Faradaic efficiency of 99.5%, and NH3 selectivity of 100%. Characterizations and theoretical calculations unveiled that the Cu-defects and OVs-Cu2O heterojunction boosted the H* yield, suppressed the hydrogen evolution reaction (HER), and served as dual reaction sites to coherently match the tandem reactions kinetics of NO3-to-NO2 and NO2-to-NH3. An integrated system was further built to combine wet flue gas denitrification and desulfurization, simultaneously converting NO and SO2 to produce the (NH4)2SO4 fertilizer. This study offers new insights into the application of low-cost Cu-based cathode for electrochemically driven wet denitrification wastewater valorization

Effects of SIRT1 activator SRT1720 on the expression and/or release of SIRT1, IL-1β and TNF-α in RAW264.7 cell culture treated with 30 mM high glucose.

<p>(A-D) RAW264.7 cells were pretreated with 1 μM SRT1720 for 6h and then cultured in 30 mM D-glucose-containing medium for additional 8h. Cells were then harvested and used for western blot analysis for SIRT1protein level (A, <i>df</i> = 5, <i>f</i> = 53.307, <i>p</i> < 0.001), and real-time PCR analysis for <i>SIRT1</i> (B, <i>df</i> = 5, <i>f</i> = 16.042, <i>p</i> = 0.002), <i>IL-1β</i> (C, <i>df</i> = 5, <i>f</i> = 328.474, <i>p</i> < 0.001) or <i>TNF-α</i> mRNA level (D, <i>df</i> = 5, <i>f</i> = 43.581, <i>p</i> < 0.001). Each data point is the mean ± SEM of <i>n</i> = 3 and normalized against corresponding β-ACTIN protien level or <i>GAPDH</i> mRNA level with the value in NG without SRT1720 group arbitrarily set as 1. (E-F) The culture medium was also collected and used for ELISA for detecting the release of IL-1β (E, <i>df</i> = 5, <i>f</i> = 5739.982, <i>p</i> < 0.001) and TNF-α (F, <i>df</i> = 5, <i>f</i> = 108.365, <i>p</i> < 0.001). Each data point is the mean ± SEM of <i>n</i> = 3. *<i>p</i> < 0.05; ***<i>p</i> < 0.001. 30 mM of D-mannitol served as an osmotic control. ‘NG’, 5.6 mM normal glucose; ‘HG’, 30 mM high glucose; ‘DM’, 30 mM D-mannitol.</p

Assessment of the mRNA level and the release of IL-1β and TNF-α in RAW264.7 cell culture treated with 30 mM high glucose over time.

<p>(A-B) RAW264.7 cells were exposed to 30 mM D-glucose and cultured for indicated time periods of 0, 4, 8, 24, 36 and 48h. The mRNA levels of two inflammatory cytokines, IL-1β (A, <i>df</i> = 5, <i>f</i> = 62.812, <i>p</i> < 0.001) and TNF-α (B, <i>df</i> = 5, <i>f</i> = 5.492, <i>p</i> = 0.03), were assessed by real-time PCR. Each data point is the mean ± SEM of <i>n</i> = 3 and normalized against corresponding <i>GAPDH</i> mRNA level with the value at 0h arbitrarily set as 1. (C-D) The release of IL-1β (C, <i>df</i> = 5, <i>f</i> = 27.324, <i>p</i> < 0.001) and TNF-α (D, <i>df</i> = 5, <i>f</i> = 48.74, <i>p</i> < 0.001) in culture medium was assessed by ELISA. Each data point is the mean ± SEM of <i>n</i> = 3. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001.</p

Effects of SIRT1 inhibitor EX527 on the expression and/or release of SIRT1, IL-1β and TNF-α in RAW264.7 cell culture treated with 30 mM high glucose.

<p>(A-D) RAW264.7 cells were pretreated with 10 μM EX527 for 6h and then cultured in 30 mM D-glucose-containing medium for additional 8h. Cells were then harvested and used for western blot analysis for SIRT1 protein level (A, <i>df</i> = 5, <i>f</i> = 6.194, <i>p</i> = 0.023), and real-time PCR analysis for <i>SIRT1</i> (B, <i>df</i> = 5, <i>f</i> = 7.118, <i>p</i> = 0.003), <i>IL-1β</i> (C, <i>df</i> = 5, <i>f</i> = 71.26, <i>p</i> < 0.001) or <i>TNF-</i>α mRNA level (D, <i>df</i> = 5, <i>f</i> = 50.638, <i>p</i> < 0.001). Each data point is the mean ± SEM of <i>n</i> = 3 and normalized against corresponding β-ACTIN protein level or <i>GAPDH</i> mRNA level with the value in NG without EX527 group arbitrarily set as 1. (E-F) The culture medium was also collected and used for ELISA for detecting the release of IL-1β (E, <i>df</i> = 5, <i>f</i> = 6057.757, <i>p</i> < 0.001) and TNF-α (F, <i>df</i> = 5, <i>f</i> = 47.79, <i>p</i> < 0.001). Each data point is the mean ± SEM of <i>n</i> = 3. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001. 30 mM of D-mannitol served as an osmotic control. ‘NG’, 5.6 mM normal glucose; ‘HG’, 30 mM high glucose; ‘DM’, 30 mM D-mannitol.</p

Assessment of the protein and mRNA level changes of SIRT1 in RAW264.7 cells treated with 30 mM high glucose over time.

<p>(A-B) Immunoblotting (A) or quantatitive RT-PCR (B) assessing the change in SIRT1 protein or mRNA level after treatment of RAW264.7 cells with 30 mM high glucose at indicated time points (0, 4, 8, 24, 36 and 48h), respectively. Each data point is the mean ± SEM of <i>n</i> = 3 and normalized against corresponding β-ACTIN protein level (A, <i>df</i> = 5, <i>f</i> = 134.651, <i>p</i> < 0.001) or <i>GAPDH</i> mRNA level (B, <i>df</i> = 5, <i>f</i> = 127.339, <i>p</i> < 0.001) with the value at 0h arbitrarily set as 1. *<i>p</i> < 0.05; **<i>p</i> < 0.01. (C) The concentration of D-glucose in the culture medium was measured over time using ONETOUCH Ultra at indicated time points (0, 4, 8, 24, 36 and 48h). Each data point is the mean ± SEM of <i>n</i> = 3.</p

Effects of SIRT1 knockdown by RNAi on the mRNA level and the release of IL-1β and TNF-α in RAW264.7 cell culture treated with 30 mM high glucose.

<p>(A) Cells cultured in NG medium were transfected with 33 nM scramble or SIRT1-specific siRNA duplex (serial number: 2195, 1003, and 576) for 6h, incubated for another 42h, and then harvested for the detection of SIRT1 suppression by western blot (<i>df</i> = 3, <i>f</i> = 38.663, <i>p</i> = 0.002). (B-C) Cells receiving RNAi treatment were continued to be cultured in 30 mM D-glucose-containing medium for additional 8h, mRNA levels of IL-1β (B, <i>df</i> = 4, <i>f</i> = 250.541, <i>p</i> < 0.001) and TNF-α (C, <i>df</i> = 4, <i>f</i> = 24.145, <i>p</i> < 0.001) were assessed by real-time PCR. Each data point is the mean ± SEM of <i>n</i> = 3 and normalized against corresponding β-ACTIN protein level (A) or <i>GAPDH</i> mRNA level (B-C) with the value in NG with scramble siRNA group arbitrarily set as 1. (D-E) The culture medium was also collected and used for ELISA for detecting the release of IL-1β (D, <i>df</i> = 4, <i>f</i> = 110.68, <i>p</i> < 0.001) and TNF-α (E, <i>df</i> = 4, <i>f</i> = 80.041, <i>p</i> < 0.001). Each data point is the mean ± SEM of <i>n</i> = 3. *<i>p</i> < 0.05; **<i>p</i> < 0.01; ***<i>p</i> < 0.001. 30 mM of D-mannitol served as an osmotic control. ‘NG’, 5.6 mM normal glucose; ‘HG’, 30 mM high glucose; ‘DM’, 30 mM D-mannitol.</p