p73 (bethyl)  (Bethyl)

Journal: The Journal of investigative dermatology

Article Title: The Robust Tumoricidal Effects of Combined BET/HDAC Inhibition in Cutaneous T-Cell Lymphoma Can Be Reproduced by ΔNp73 Depletion.

doi: 10.1016/j.jid.2022.06.005

Figure Lengend Snippet: Figure 4. TAp73/DNp73 protein ratio is increased at 72 hours by BETi/HDACi in CTCL lines and leukemic Se´zary tumor cells but not in normal CD4D T cells. (a) Shifts in TAp73/DNp73 protein ratios detected by flow cytometry in CTCL lines, SS, and normal CD4þ T cells. *P < 0.05 and **P < 0.01. Graphs show the mean SEM for CTCL lines and CD4þ T cells and mean SD for Se´zary cells. (b) Immunoblots show absolute increases in TAp73 and decreases in DNp73 among CTCL lines. BETi, BET inhibitor; CTCL, cutaneous T-cell lymphoma; h, hour; HDACi, histone deacetylase inhibitor; SS, Se´zary syndrome; Veh, vehicle.

Article Snippet: Antibody List Target Source Catalog Number Application Dilution BIM Cell Signaling Technology 2933 Immunoblot 1:1,000 BAX Cell Signaling Technology 5023 Immunoblot 1:1,000 BCL2 Cell Signaling Technology 4223 Immunoblot 1:1,000 b-actin Sigma-Aldrich A2228 Immunoblot 1:5,000 DNp73 Novus Biological NBP2-24873 Immunoblot, flow cytometry, IHC 1:1,000, 1:200, 1:50 TAp73 Bethyl Laboratory A300-126A Immunoblot 1:1,000 TAp73 GeneTex GTX46123 Flow cytometry, IHC 1:200, 1:50 CD3 Abcam ab16669 IHC 1:150 H3K27ac Active Motif 39133 ChIP 10 mg per ChIP RNA Pol II Active Motif 39097 ChIP 10 mg per ChIP Rabbit IgG Abcam ab37415 ChIP 10 mg per ChIP Abbreviations: ChIP, chromatin immunoprecipitation; IHC, immunohistochemistry; Pol, polymerase. www.jidonline.org 3261.e4

Techniques: Cytometry, Western Blot, Histone Deacetylase Assay

Journal: The Journal of investigative dermatology

Article Title: The Robust Tumoricidal Effects of Combined BET/HDAC Inhibition in Cutaneous T-Cell Lymphoma Can Be Reproduced by ΔNp73 Depletion.

doi: 10.1016/j.jid.2022.06.005

Figure Lengend Snippet: Figure 3. TP73 transcript alterations in CTCL lines induced by BETi/HDACi. (a) Heat map of row Z-scores for DNp73 transcript alterations over time. (b) Volcano plot of aggregated CTCL cell line data 96 hours after BETi/HDACi. DNp73 transcript (circled) decreased 18.9 fold (P < 0.005). DNp73 was also significantly decreased at 6 hours (2.4-fold; P < 0.04) and 48 hours (4.9-fold; P < 0.02) (not shown). (c) Quantitative RT-PCR of TAp73 and DNp73 over time. *P < 0.05 and **P < 0.01. Triplicate values for all CTCL line data are shown in a‒c. BETi, BET inhibitor; CTCL, cutaneous T-cell lymphoma; h, hour; HDACi, histone deacetylase inhibitor.

Article Snippet: Antibody List Target Source Catalog Number Application Dilution BIM Cell Signaling Technology 2933 Immunoblot 1:1,000 BAX Cell Signaling Technology 5023 Immunoblot 1:1,000 BCL2 Cell Signaling Technology 4223 Immunoblot 1:1,000 b-actin Sigma-Aldrich A2228 Immunoblot 1:5,000 DNp73 Novus Biological NBP2-24873 Immunoblot, flow cytometry, IHC 1:1,000, 1:200, 1:50 TAp73 Bethyl Laboratory A300-126A Immunoblot 1:1,000 TAp73 GeneTex GTX46123 Flow cytometry, IHC 1:200, 1:50 CD3 Abcam ab16669 IHC 1:150 H3K27ac Active Motif 39133 ChIP 10 mg per ChIP RNA Pol II Active Motif 39097 ChIP 10 mg per ChIP Rabbit IgG Abcam ab37415 ChIP 10 mg per ChIP Abbreviations: ChIP, chromatin immunoprecipitation; IHC, immunohistochemistry; Pol, polymerase. www.jidonline.org 3261.e4

Techniques: Quantitative RT-PCR, Histone Deacetylase Assay