Journal: Cell death discovery

Article Title: ABCB1 confers resistance to carboplatin by accumulating stem-like cells in the G2/M phase of the cell cycle in p53 null ovarian cancer.

doi: 10.1038/s41420-025-02435-7

Figure Lengend Snippet: Fig. 5 Reversal of the resistant phenotype conferred by upregulated ABCB1 through ΔNp73 suppression. A Quantitative gene expression of ΔNp73 in nonR and carR-SKOV3 cells (p = 0.0005). B Immunoblotting analysis of ΔNp73 protein levels in nonR and carR-SKOV3 cells via immunoblotting assay. Validation of ΔNp73 silencing in carR-SKOV3 compared to non-treated and negative control (NC) transfected-groups both at protein (C) and mRNA (D) levels (non- vs NC-transfected; p = 0.351, non- vs siΔNp73-transfected; p = 0.00002). E Quantitative gene expression of ABCB1 in carR-SKOV3 (non-treated, siNC-transfected, and siΔNp73-transfected) cells (non-treated vs siNC-transfected; p = 0.468, non-treated vs siΔNp73-transfected; p = 0.00005). F Relative cell growth rates of siΔNp73-transfected carR-SKOV3 cells compared to non- transfected and NC-transfected cells (24 h: non-treated vs siNC-transfected; p = 0.017, 48 h: non-treated vs siNC-transfected; p = 0.0002). G Comparison of Ki67 expression in carR-SKOV3 (non-treated, siNC-transfected, and siΔNp73-transfected) cells (non-treated vs siNC- transfected; p = 0.013). H Cell viability assay in carR-SKOV3 (non-treated, siNC-transfected, and siΔNp73-transfected) compared to nonR- SKOV3 after induction of additional damage up to 72 h. Data represent the means ± SD from triplicate experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, NS not significant.

Article Snippet: After blocking procedure, membranes were incubated with primary antibodies against p53 (7F5), phospho-p53 (E9Y4U; Ser15), PARP1 (46D11), cleaved-PARP1 (D64E10; Asp214), BCL-XL (54H6), phospho-cdc25c (63F9; Ser216), cyclin B1 (D5C10), ΔNp73 (38C674.2; Novus, USA), and β-actin (all from Cell signaling, USA), respectively, and finally visualized by ECL solution (Thermo, USA).

Techniques: Gene Expression, Western Blot, Biomarker Discovery, Negative Control, Transfection, Comparison, Expressing, Viability Assay

Journal: Blood Cancer Discovery

Article Title: HTLV-1 bZIP Factor-Induced Reprogramming of Lactate Metabolism and Epigenetic Status Promote Leukemic Cell Expansion

doi: 10.1158/2643-3230.bcd-22-0139

Figure Lengend Snippet: Figure 1. HBZ protein and RNA both upregulate TP73. A, Schematic diagram for retroviral transfer of HBZ into primary mouse CD4+ T cells. Each construct encodes WT HBZ, HBZ RNA (ATG is converted to TTG) or HBZ protein (SM, silent mutations; ref. 5). Created with BioRender.com. B, A heat map of the top 500 differentially expressed genes in HBZ WT and its mutants compared with the vector, calculated from RNA-seq data. C, Shared transcrip- tomes (left; the number of genes) and open chromatin regions (right) associated with HBZ WT and its mutants are shown in Venn diagrams. D, Volcano plots of differentially expressed genes. Fold change and adjusted P value (Padj) are plotted for genes that are upreulgated (yellow) or downregulated (blue) compared with the vector. E, TP73 gene maps (upper, mouse; lower, human) depicting the major two isoforms, TAp73 and DNp73, and their promot- ers. (continued on next page)

Article Snippet: Immunoblotting was conducted with antibodies against HBZ described above, Flag (Sigma-Aldrich, #F7425), HA (MBL International, #561-5), α-Tubulin (Sigma-Aldrich, #T6199), EZH2 (Cell Signaling Technology, #5246S), TAp73 (Abcam, #ab14430), and DNp73 (Novus Biologicals, #NBP224873).

Techniques: Retroviral, Construct, Plasmid Preparation, RNA Sequencing

Journal: Blood Cancer Discovery

Article Title: HTLV-1 bZIP Factor-Induced Reprogramming of Lactate Metabolism and Epigenetic Status Promote Leukemic Cell Expansion

doi: 10.1158/2643-3230.bcd-22-0139

Figure Lengend Snippet: Figure 1. (Continued) F, Transcripts per million (TPM) of TAp73 in the transduced cells (n = 3). G, A volcano plot resulting from RNA-seq that compares the CD4+ T cells of HBZ-Tg mice with those of WT mice (n = 3). H, TPM of TAp73 in HBZ-Tg and WT mouse CD4+ T cells (n = 3). I–K, TP73 expression in ATL. A volcano plot resulting from RNA-seq compares CD4+ T cells of ATL patients (n = 7) with those of healthy donors (n = 10; I). mRNA expression of TAp73 (J) and DNp73 (K) by RT-qPCR in CD4+ T cells of ATL patients (acute type, n = 28; lymphoma type, n = 5; chronic type, n = 7) and healthy donors (hCD4; n = 8). Results are plotted as mean ± SD, using one-way ANOVA followed by the post hoc Steel test (J and K). ***, P < 0.001; ns, not significant.

Article Snippet: Immunoblotting was conducted with antibodies against HBZ described above, Flag (Sigma-Aldrich, #F7425), HA (MBL International, #561-5), α-Tubulin (Sigma-Aldrich, #T6199), EZH2 (Cell Signaling Technology, #5246S), TAp73 (Abcam, #ab14430), and DNp73 (Novus Biologicals, #NBP224873).

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR

Journal: Blood Cancer Discovery

Article Title: HTLV-1 bZIP Factor-Induced Reprogramming of Lactate Metabolism and Epigenetic Status Promote Leukemic Cell Expansion

doi: 10.1158/2643-3230.bcd-22-0139

Figure Lengend Snippet: Figure 3. HBZ RNA activates both TAp73 and DNp73 promoters via BATF3-IRF4 transcriptional machinery. A, Transcripts per million (TPM) of BATF3 in HBZ-transduced murine CD4+ T cells (n = 3). B, The top-ranked enriched motifs of HBZ RNA-transduced murine CD4+ T cells with their log2 P values from the findMotifsGenome results (HOMER). C, Chromatin accessibility (ATAC-seq), H3K27ac enrichment, BATF3/IRF4 binding regions (ChIP-seq; SRX2548278 and SRX2548284; ref. 25) analyzed by using the ChIP-Atlas (73), and transcripts (RNA-seq) of the TP73 gene in HTLV-1–negative or –positive human T-cell lines. D, Promoter assays of the HTLV-1–specific open region identified in C (hg19 genome region of chr1:3593076–3594185) with a minimal promoter (minP) in HEK293 cells cotransfected with WT or mutant HBZ. The open region was inserted into pNL3.2.CMV after cloning of the genome region as shown in a schematic of construct. E and F, The IRF4/AP-1 motifs identified within the open region (hg19 genome region of chr1:3593076–3594185) were subjected to promoter assays with IRF4 and/or BATF3 induction: for the promoter of TAp73 (E) and the promoter of DNp73 (F; n = 3). A schematic of the assay construct is shown above the corresponding bar plot. Results are plotted with mean ± SD, using one-way ANOVA with the post hoc Dunnet test (D–F). *, P < 0.05; ***, P < 0.001; ns, not significant.

Article Snippet: Immunoblotting was conducted with antibodies against HBZ described above, Flag (Sigma-Aldrich, #F7425), HA (MBL International, #561-5), α-Tubulin (Sigma-Aldrich, #T6199), EZH2 (Cell Signaling Technology, #5246S), TAp73 (Abcam, #ab14430), and DNp73 (Novus Biologicals, #NBP224873).

Techniques: Binding Assay, ChIP-sequencing, RNA Sequencing, Mutagenesis, Cloning, Construct

Journal: Blood Cancer Discovery

Article Title: HTLV-1 bZIP Factor-Induced Reprogramming of Lactate Metabolism and Epigenetic Status Promote Leukemic Cell Expansion

doi: 10.1158/2643-3230.bcd-22-0139

Figure Lengend Snippet: Figure 5. TAp73 inactivation causes ATL cell death and intracellular acidification due to lactate accumulation. A, Mating diagram of HBZ-Tg mice with TP73 knockout mice (TAp73−/− or DNp73−/−) created by CRISPR/Cas9. B, Cumulative incidence of skin inflammation in HBZ-Tg mice (n = 32) compared with HBZ-Tg/TAp73−/− mice (left; n = 18) or HBZ-Tg/DNp73−/− mice (right; n = 11). The P values determined by the Gray test are shown. C and D, Changes in metabolites between HBZ-Tg and HBZ-Tg/TAp73−/− mouse CD4+ T cells. Shown are the top 25 differentially altered metabolites with the normalized area under the curve (AUC; mean-centered and divided by the standard deviation of each variable; C) and enriched metabolite sets (D), both of which were analyzed by using the MetaboAnalyst (78). E and F, Intracellular lactate (E) and pyruvate (F) levels in the murine CD4+ T cells (n = 3). Calculated ion intensi- ties (normalized AUC) are shown. (continued on following page)

Article Snippet: Immunoblotting was conducted with antibodies against HBZ described above, Flag (Sigma-Aldrich, #F7425), HA (MBL International, #561-5), α-Tubulin (Sigma-Aldrich, #T6199), EZH2 (Cell Signaling Technology, #5246S), TAp73 (Abcam, #ab14430), and DNp73 (Novus Biologicals, #NBP224873).

Techniques: Knock-Out, CRISPR, Standard Deviation

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