Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A: Effect of different catechin (Cat) concentrations on viable cell density (VCD) (blue) and viability (yellow) B : effect of different catechin concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different epicatechin (EC) concentrations on viable cell density (VCD) (blue) and viability (yellow). B : effect of different epicatechin concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different gallocatechin gallate (GCG) concentrations on viable cell density (VCD) (blue) and viability (yellow). B : effect of different GCG concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A: Effect of different epigallocatechin gallate (EGCG) concentrations on viable cell density (VCD) (blue) and viability (yellow) B : effect of different EGCG concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A: Effect of different kaempferol (Kmp) concentrations on viable cell density (VCD) (blue) and viability (yellow) B : effect of different kaempferol concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different luteolin (Ltl) concentrations on viable cell density (VCD) (blue) and viability (yellow) B : effect of different luteolin concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different curcumin (Crc) concentrations on viable cell density (VCD) (blue) and viability (yellow). B : effect of different curcumin concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different caffeic acid phenethyl ester (CAPE) concentrations on viable cell density (VCD) (blue) and viability (yellow). B : effect of different CAPE concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different resveratrol (Rsv) concentrations on viable cell density (VCD) (blue) and viability (yellow) B : effect of different resveratrol concentrations (10-100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different piceid (Pcd) concentrations on viable cell density (VCD) (blue) and viability (yellow). B : effect of different piceid concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Antioxidants

Article Title: Screening Naturally Occurring Phenolic Antioxidants for Their Suitability as Additives to CHO Cell Culture Media Used to Produce Monoclonal Antibodies

doi: 10.3390/antiox8060159

Figure Lengend Snippet: A : Effect of different α-tocopherol (Tcp) concentrations on viable cell density (VCD) (blue) and viability (yellow) B : effect of different α-tocopherol concentrations (10–100 µM) IgG concentration (grey) and relative IgG production (qP) (orange) for CHO cells grown in 24-well plates. Each data point represents the average from a quadruplicate, and the error bars correspond to ±SD. * p -value < 0.05 for the treatment compared to the control.

Article Snippet: The cell line used was a stable recombinant DG44 CHO cell line that expressed an IgG monoclonal antibody, provided by Fujifilm Diosynth.

Techniques: Concentration Assay

Journal: Molecular cell

Article Title: Co-condensation between transcription factor and coactivator p300 modulates transcriptional bursting kinetics.

doi: 10.1016/j.molcel.2021.01.031

Figure Lengend Snippet: Figure 1. Transcriptional coactivator p300 forms liquid-like clusters that display spatial correlations with TFs (A) Representative live-cell image of endogenously tagged mEGFP-p300, and representative immunofluorescence (IF) image of p300 in mESC. (B) FRAP experiments on p300 foci in U2OS cells with exogenously expressed mEGFP-p300. Foci were photobleached at t = 0 s. Error bars indicate ± SEM (n = 11). (C) Representative live-cell images of U2OS cells cotransfected with mEGFP-p300 and mCherry-CBP (left) and FRAP experiments on mEGFP-CBP foci (right). Error bars indicate ± SEM (n = 11). (D) Distribution of p300 foci in individual mESCs. (E) Number of p300 foci in a mESC undergoing mitosis (Figure S1K). (F) Trajectory lifetime distribution of identified p300 foci from 41 mESCs. (G) (Top) Average fluorescence signals of p65 and p300 of 288 p65 foci in an SK-N-AS cell (Figure S2B). (Bottom) Population-averaged p65 and p300 signals under TNF-a treatment. Error bars indicate ± SEM. (H) Analysis analogous to that in (G) for cells without TNF-a treatment.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies mouse monoclonal anti-p300 Santa Cruz Biotechnology Cat# sc-32244; RRID: AB_628076 Rabbit monoclonal anti-p300 Cell Signaling Technology Cat# 86377; RRID: AB_2800077 Goat Anti-Rabbit IgG H&L (Alexa Fluor 594) antibody Abcam Cat# ab150080; RRID:AB_2650602 Goat Anti-Mouse IgG H&L (Alexa Fluor 488) antibody Abcam Cat# ab150113; RRID:AB_2576208 Mouse monoclonal NFkappaB p65 (F-6) antibody Santa Cruz Biotechnology Cat# sc-8008; RRID:AB_628017 Rabbit monoclonal anti-Sox2 Cell Signaling Technology Cat# 23064; RRID:AB_2714146 Rabbit polyclonal anti-p300 (Lys1499) antibody Cell Signaling Technology Cat# 4771; RRID:AB_2262406 Rabbit polyclonal anti-histone H3K27ac antibody Active Motif Cat# 39133; RRID:AB_2561016 Anti-rabbit IgG, HRP-linked Antibody Cell Signaling Technology Cat# 7074; RRID:AB_2099233 Bacterial and virus strains E. coli, Trelief 5a Chemically Competent Cell TSINGKE Cat# TSC01 E.coli, DH5a Competent Cell CWBIO Cat# CW0808 Chemicals, peptides, and recombinant proteins bisBenzimide H33342 trihydrochloride SIGMA Cat# B2261 Doxycycline Clontech Laboratories Cat# 8634-1 (+/ )-JQ1 SIGMA Cat# SML0974 Lipofectamine LTX Reagent with PLUS Reagent Thermo Fisher Scientific Cat# 15338100 Leukemia Inhibitory Factor (LIF) ESGRO ESG1107 Trichostatin A (TSA) Selleck Cat# S1045 A-485 Selleck Cat# S8740 mEGFP This study N/A mEGFP-p300CTR This study N/A mEGFP-p300CTR (Q2A) This study N/A Deposited data Imaging data This study, Mendeley Data https://dx.doi.org/10.17632/bnm84ghmvx.1 mESC wild type RNA-seq This study GEO: GSE161958 mESC mEGFP-p300-tagged cell line RNA-seq This study GEO: GSE161958 Experimental models: cell lines Mouse: E14 cells (E14Tg2a.4) Michael B. Elowitz Lab N/A mEGFP-p300-tagged E14 cells (Clone W9) This study N/A Human: U2OS ATCC N/A Human: U2OS MLCPP7-p53TAD This study N/A Human: U2OS MLCPP7-p65TAD This study N/A Human: U2OS MLCPP7-p53TAD-p300CTR This study N/A Human: U2OS MLCPP7-p53TAD-p300CTR (Q2A) This study N/A Human: U2OS MLCPP7-TD-p53TAD This study N/A Human:HEK293T cells ATCC N/A Human: SK-N-AS Ping Wei Lab N/A Chinese hamsters:A03_1 CHO DG44 cells Guohong Li Lab N/A Recombinant DNA CDS:CRY2 Ping Wei Lab N/A CDS:p300,human Mailgene biosciences N/A (Continued on next page) e1 Molecular Cell 81, 1682–1697.e1–e7, April 15, 2021

Techniques:

Journal: Molecular cell

Article Title: Co-condensation between transcription factor and coactivator p300 modulates transcriptional bursting kinetics.

doi: 10.1016/j.molcel.2021.01.031

Figure Lengend Snippet: Figure 2. TF/p300 co-condensation can be initiated by self-association of the transactivation domains (TADs) of TFs and enhances the sta- bility of TF clusters (A) Assay schematic. (B and C) Snapshots of representative mEGFP-p300 mESCs expressing Cry2-mCherry-p65TAD (B) or Cry2-mCherry-p53TAD (C) before and after blue light stimulation. See Figure S3D for the indicated region in (B). (D) Assay schematic. (E) Snapshots of a representative U2OS cell expressing Cry2-mCherry-p65TAD before and after light stimulation. This cell contains exogenously expressed mEGFP-p300 (i.e., p300 overexpression). Cluster formation after stimulation was visually quantified across a range of nuclear mCherry levels for cells without (top, n = 173 cells) or with (bottom, n = 141 cells) p300 overexpression. 1 indicates observable condensation, and 0 indicates no observable condensation. Dashed lines indicate potential bistable regions. (F) Assay analogous to that in (E) for Cry2-mCherry-p53TAD. n = 152 (top), 100 (bottom) cells.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies mouse monoclonal anti-p300 Santa Cruz Biotechnology Cat# sc-32244; RRID: AB_628076 Rabbit monoclonal anti-p300 Cell Signaling Technology Cat# 86377; RRID: AB_2800077 Goat Anti-Rabbit IgG H&L (Alexa Fluor 594) antibody Abcam Cat# ab150080; RRID:AB_2650602 Goat Anti-Mouse IgG H&L (Alexa Fluor 488) antibody Abcam Cat# ab150113; RRID:AB_2576208 Mouse monoclonal NFkappaB p65 (F-6) antibody Santa Cruz Biotechnology Cat# sc-8008; RRID:AB_628017 Rabbit monoclonal anti-Sox2 Cell Signaling Technology Cat# 23064; RRID:AB_2714146 Rabbit polyclonal anti-p300 (Lys1499) antibody Cell Signaling Technology Cat# 4771; RRID:AB_2262406 Rabbit polyclonal anti-histone H3K27ac antibody Active Motif Cat# 39133; RRID:AB_2561016 Anti-rabbit IgG, HRP-linked Antibody Cell Signaling Technology Cat# 7074; RRID:AB_2099233 Bacterial and virus strains E. coli, Trelief 5a Chemically Competent Cell TSINGKE Cat# TSC01 E.coli, DH5a Competent Cell CWBIO Cat# CW0808 Chemicals, peptides, and recombinant proteins bisBenzimide H33342 trihydrochloride SIGMA Cat# B2261 Doxycycline Clontech Laboratories Cat# 8634-1 (+/ )-JQ1 SIGMA Cat# SML0974 Lipofectamine LTX Reagent with PLUS Reagent Thermo Fisher Scientific Cat# 15338100 Leukemia Inhibitory Factor (LIF) ESGRO ESG1107 Trichostatin A (TSA) Selleck Cat# S1045 A-485 Selleck Cat# S8740 mEGFP This study N/A mEGFP-p300CTR This study N/A mEGFP-p300CTR (Q2A) This study N/A Deposited data Imaging data This study, Mendeley Data https://dx.doi.org/10.17632/bnm84ghmvx.1 mESC wild type RNA-seq This study GEO: GSE161958 mESC mEGFP-p300-tagged cell line RNA-seq This study GEO: GSE161958 Experimental models: cell lines Mouse: E14 cells (E14Tg2a.4) Michael B. Elowitz Lab N/A mEGFP-p300-tagged E14 cells (Clone W9) This study N/A Human: U2OS ATCC N/A Human: U2OS MLCPP7-p53TAD This study N/A Human: U2OS MLCPP7-p65TAD This study N/A Human: U2OS MLCPP7-p53TAD-p300CTR This study N/A Human: U2OS MLCPP7-p53TAD-p300CTR (Q2A) This study N/A Human: U2OS MLCPP7-TD-p53TAD This study N/A Human:HEK293T cells ATCC N/A Human: SK-N-AS Ping Wei Lab N/A Chinese hamsters:A03_1 CHO DG44 cells Guohong Li Lab N/A Recombinant DNA CDS:CRY2 Ping Wei Lab N/A CDS:p300,human Mailgene biosciences N/A (Continued on next page) e1 Molecular Cell 81, 1682–1697.e1–e7, April 15, 2021

Techniques: Expressing, Over Expression

Journal: Molecular cell

Article Title: Co-condensation between transcription factor and coactivator p300 modulates transcriptional bursting kinetics.

doi: 10.1016/j.molcel.2021.01.031

Figure Lengend Snippet: Figure 4. TF/p300 clustering results in the transactivation of p300 in the clusters and the subsequent recruitment of Brd4 (A) Representative IF images of U2OS cells containing synthetic TF variants (stained with anti-p300 and anti-p300 K1499ac antibodies). (B) Quantification of p300 activation by TF/p300 co-condensation with western blotting. Relative intensities were calculated as ratios between p300 and K1499ac. (C and D) ChIP-qPCR quantifications of H3K27ac enrichment at the promoter and gene body for genes regulated by rTetR-p65TAD or rTetR-p53TAD (with doxycycline [dox], n = 3). (E and F) Snapshots of representative U2OS cells containing rTetR-iRFP-p65TAD and mEGFP-p300 under DMSO (E) or A-485 treatment (F), and quantifications of the numbers and areas of foci at the indicated time points (using ImageJ). Treatments started at 0 min. No dox was added. (G) Assay schematic for (H)–(K).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies mouse monoclonal anti-p300 Santa Cruz Biotechnology Cat# sc-32244; RRID: AB_628076 Rabbit monoclonal anti-p300 Cell Signaling Technology Cat# 86377; RRID: AB_2800077 Goat Anti-Rabbit IgG H&L (Alexa Fluor 594) antibody Abcam Cat# ab150080; RRID:AB_2650602 Goat Anti-Mouse IgG H&L (Alexa Fluor 488) antibody Abcam Cat# ab150113; RRID:AB_2576208 Mouse monoclonal NFkappaB p65 (F-6) antibody Santa Cruz Biotechnology Cat# sc-8008; RRID:AB_628017 Rabbit monoclonal anti-Sox2 Cell Signaling Technology Cat# 23064; RRID:AB_2714146 Rabbit polyclonal anti-p300 (Lys1499) antibody Cell Signaling Technology Cat# 4771; RRID:AB_2262406 Rabbit polyclonal anti-histone H3K27ac antibody Active Motif Cat# 39133; RRID:AB_2561016 Anti-rabbit IgG, HRP-linked Antibody Cell Signaling Technology Cat# 7074; RRID:AB_2099233 Bacterial and virus strains E. coli, Trelief 5a Chemically Competent Cell TSINGKE Cat# TSC01 E.coli, DH5a Competent Cell CWBIO Cat# CW0808 Chemicals, peptides, and recombinant proteins bisBenzimide H33342 trihydrochloride SIGMA Cat# B2261 Doxycycline Clontech Laboratories Cat# 8634-1 (+/ )-JQ1 SIGMA Cat# SML0974 Lipofectamine LTX Reagent with PLUS Reagent Thermo Fisher Scientific Cat# 15338100 Leukemia Inhibitory Factor (LIF) ESGRO ESG1107 Trichostatin A (TSA) Selleck Cat# S1045 A-485 Selleck Cat# S8740 mEGFP This study N/A mEGFP-p300CTR This study N/A mEGFP-p300CTR (Q2A) This study N/A Deposited data Imaging data This study, Mendeley Data https://dx.doi.org/10.17632/bnm84ghmvx.1 mESC wild type RNA-seq This study GEO: GSE161958 mESC mEGFP-p300-tagged cell line RNA-seq This study GEO: GSE161958 Experimental models: cell lines Mouse: E14 cells (E14Tg2a.4) Michael B. Elowitz Lab N/A mEGFP-p300-tagged E14 cells (Clone W9) This study N/A Human: U2OS ATCC N/A Human: U2OS MLCPP7-p53TAD This study N/A Human: U2OS MLCPP7-p65TAD This study N/A Human: U2OS MLCPP7-p53TAD-p300CTR This study N/A Human: U2OS MLCPP7-p53TAD-p300CTR (Q2A) This study N/A Human: U2OS MLCPP7-TD-p53TAD This study N/A Human:HEK293T cells ATCC N/A Human: SK-N-AS Ping Wei Lab N/A Chinese hamsters:A03_1 CHO DG44 cells Guohong Li Lab N/A Recombinant DNA CDS:CRY2 Ping Wei Lab N/A CDS:p300,human Mailgene biosciences N/A (Continued on next page) e1 Molecular Cell 81, 1682–1697.e1–e7, April 15, 2021

Techniques: Staining, Activation Assay, Western Blot, ChIP-qPCR