Journal: Reproduction

Article Title: Increased rounds of gonadotropin stimulation have side effects on mouse fallopian tubes and oocytes

doi: 10.1530/rep-17-0687

Figure Lengend Snippet: Figure 2. Western blot analysis of p53 and p-p53 in fallopian tubes (FT) Ctr: Control mice; 6R and 8R: mice undergoing 6 rounds and 8 rounds of gonadotropin stimulation (A). Data

Article Snippet: Mouse monoclonal 89 p53 and OCT3/4, rabbit polyclonal Cyclin D1, phospho-B-catenin (Thr41/Ser45), B-catenin, 90 5 phospho-AKT (Ser473), AKT, phospho-GSK3B (Ser9), GSK3B and SOX2 primary 91 antibodies were purchased from Santa Cruz Biotechnology (CA, USA); mouse monoclonal 92 phospho-p53 (Ser15), rabbit monoclonal phospho-CHK1 (Ser345) and phospho-H2A.X 93 (Ser139) were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Western Blot, Control

Journal: Cells

Article Title: Towards Understanding the Development of Breast Cancer: The Role of RhoJ in the Obesity Microenvironment

doi: 10.3390/cells13020174

Figure Lengend Snippet: Different antibodies used and their dilutions.

Article Snippet: Phospho-p53 (S15) , 1:1000 , Rabbit , Cell signaling, Danvers, MA, USA , 9286s.

Techniques:

Journal: Scientific Reports

Article Title: Regulation of cardiomyocyte DNA damage and cell death by the type 2A protein phosphatase regulatory protein alpha4

doi: 10.1038/s41598-021-85616-5

Figure Lengend Snippet: Regulation of the DNA damage response by α4 protein expression in cardiomyocytes. H9c2 cardiomyocytes were transfected with either 50 nM non-targeting control (NTC) siRNA or 50 nM siRNA specific to rat α4 protein (siα4) for 1–4 days (n = 4) and the KD of α4 protein expression was confirmed by Western analysis using an antibody raised against human α4 ( a ). α4 protein expression was knocked down for 4 days and Western analysis (short and long exposures) was used to probe samples (n = 3) with an antibody specific to phosphorylated S / T QG motifs within ATM/ATR substrates. Arrows indicate substrates whose phosphorylation status (n = 3) was elevated in response to α4 KD ( b ). The expression and phosphorylation (Ser15) of p53 were determined by Western analysis using rodent-specific antibodies raised against mouse p53 protein in samples following 4 days of α4 KD, n = 5 ( c ). The expression of PARP was determined by Western analysis (short and long exposures) following 1–4 days of α4 KD, n = 3. Arrows indicate full length PARP (116 kDa) and cleaved fragments (89 kDa and 24 kDa) ( d ). All data represents data mean values ± SEM and statistical significance vs. non-targeting control (NTC) was determined by a one-tailed Student’s T-test. Expression of β-actin was used to confirm equal protein loading between samples.

Article Snippet: Antibodies to detect Bax (#2772), Bcl-xL (#2762), phosphospecific ATM/ATR substrate (#6966), IGBP1 (α4) (#5699), PARP (#9542), phosphospecific (Ser139) H2AX (#9718), H2AX (#2595), rodent-specific p53 (#32532), rodent-specific phosphospecific (Ser15) p53 (#12571) and V5-tag (#13202) were obtained from Cell Signaling Technology (USA).

Techniques: Expressing, Transfection, Control, Western Blot, Phospho-proteomics, One-tailed Test

Journal: Nature Communications

Article Title: Timing and cell specificity of senescence drives postnatal lung development and injury

doi: 10.1038/s41467-023-35985-4

Figure Lengend Snippet: C57BL/6 J neonatal mice (<12 h old) were exposed to air or hyperoxia (>95% O 2 ) for 3 days. Some mice were then allowed to recover in room air until pnd60. a p21 mRNA was measured using qRT-PCR in mouse lungs at pnd3 and pnd7. b Immunofluorescence was performed to determine expression of p21 protein. Representative images of p21 staining (red) in mouse lungs at pnd7. Arrows denote p21 positive cells. Bar size: 25 µm. c Cells positive for p21 were counted and normalized to total numbers of nuclei in the lung. d p53 protein levels were determined by Western blot in mouse lungs at pnd7. e Histogram shows p53 protein levels by hyperoxic exposure in mouse lungs at pnd3, pnd7, pnd10 and pnd60. f Phosphorylation of p53 at serine 18 was measured by ELISA in mouse lungs at both pnd3 and pnd7. g Phosphorylation of p53 at serine 18 was measured by ELISA in C 12 FDG sorted lung cells at pnd7. h p16 mRNA was measured using qRT-PCR in mouse lungs at pnd3, pnd7, pnd10 and pnd60. Data are expressed as mean ± SEM. N = 4–6 mice per group. Source data are provided as a Source Data file. † P < 0.05 vs air group at pnd3. One-way ANOVA followed by Tukey post-test was used for multiple comparisons ( a , c , e , f , h ), while t -test was used in panel ( g ). * P < 0.05, ** P < 0.01, *** P < 0.001 vs corresponding air group.

Article Snippet: Samples were loaded in duplicate onto a plate, and phosphorylation of p53 was measured using the phosphorylated Ser15 p53 ELISA kit (Cell Signaling #7365) according to manufacturer’s instructions .

Techniques: Quantitative RT-PCR, Immunofluorescence, Expressing, Staining, Western Blot, Phospho-proteomics, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Experimental Medicine

Article Title: Loss of tRNA-modifying enzyme Elp3 activates a p53-dependent antitumor checkpoint in hematopoiesis

doi: 10.1084/jem.20200662

Figure Lengend Snippet: Deficiency in Elongator activates the p53 and Atf4 stress response pathways in hematopoietic progenitors. (A) Representative pseudoblot of eIF2α and phospho-Ser51eIF2α (lower bands) protein expression on protein extracts of Elp3 fl/ f l and Elp3 HscKO MyPs obtained by Wes nanocapillary immunoassay. Vinculin was used as loading control for quantification (upper band, positive control MG132-treated BM). (B and C) MA plots of the changes in mRNA expression in Elp3 HscKO vs. Elp3 fl/fl LT-HSCs (B) and MyPs (C; left, same data as in for MyPs) and GSEAR analysis of upregulated Hallmark MSigDb gene sets (right, *, FWER P < 0.05; **, FWER P < 0.01; ****, FWER P < 1.10 −4 ). (D) Intracellular flow cytometric profile of nuclear p53 expression (left) and comparison of the p53 signal intensity (right) in Elp3 fl/fl and Elp3 HscKO HSPCs (two-way ANOVA followed by Tukey HSD tests, n = 10/group, data pooled from three experiments). (E) Flow cytometric profile of p53 signal in LT-HSCs and MyPs from Elp3 HscKO 6-Gy irradiated or Elp3/Trp53 HscDKO mice as positive and negative staining control, respectively. (F) Flow cytometric profile of phospho-Ser15-p53 expression in LSK and MyP from Elp3 fl/fl or Trp53 HscKO mice 6-Gy irradiated or not as positive and negative staining control, respectively. (G) Intracellular flow cytometric (ICFC) profile of Atf4 expression (left) and comparison of the percentages of Atf4 high cells (right) in Elp3 fl/fl and Elp3 HscKO HSPCs (****, P < 1.10 −4 in a two-way ANOVA followed by Tukey HSD tests, n = 8–9/group, data pooled from two experiments). (H) Flow cytometric profile of Atf4 expression in HSCs and MyPs from Elp3 fl/fl , Elp3 fl/fl MxCre T/+ , Atf4 HscTG , or Elp3/Atf4 fl/fl MxCre T/+ mice as positive and negative staining control, respectively. FMO, fluorescence minus one; NES, normalized enrichment score. ****, P < 1.10 −4 .

Article Snippet: ELP3 rabbit mAb (clone D5H12, cat#5728, RRID:AB_11178379), β-actin mouse mAb (clone 8H10D10, cat#3700, RRID:AB_2242334), anti-mouse IgG, HRP-linked antibody (cat#7076, RRID:AB_330924), anti-rabbit IgG, HRP-linked antibody (cat#7074, RRID:AB_2099233), ATF4 rabbit mAb (clone D4B8, cat#11815S, RRID:AB_2616025), phospho-eIF2α (Ser51) rabbit mAb (clone D9G8, cat#3398S, RRID:AB_2096481), eIF2α rabbit mAb (clone D7D3, cat#5324S, RRID:AB_10692650), p53 Alexa Fluor 647 mouse mAb (clone 1C12, cat#2533, RRID:AB_2256294), phospho-p53 (Ser15) PE mouse mAB (clone 16G8, cat#8514, RRID:AB_11178384), 4E-BP1 PE rabbit mAb (clone 53H11, cat#34470, RRID:AB_2799054), phospho-4E-BP1 (Thr37/46) Alexa Fluor 488 rabbit mAb (clone 236B4, cat#2846, RRID:AB_2292749), and c-Myc Alexa Fluor 488 rabbit mAb (clone D84C12, Cat#12855, RRID:AB_2798045) were purchased from Cell Signaling Technology.

Techniques: Expressing, Control, Positive Control, Comparison, Irradiation, Negative Staining, Fluorescence

Journal: The Journal of Experimental Medicine

Article Title: Loss of tRNA-modifying enzyme Elp3 activates a p53-dependent antitumor checkpoint in hematopoiesis

doi: 10.1084/jem.20200662

Figure Lengend Snippet: Elp3 deletion activates p53 and Atf4 stress responses in hematopoietic progenitors. (A) Principal component analysis on the transcriptome of Elp3 fl/fl and Elp3 HscKO LT-HSCs and MyPs. ( B and C ) GSEAR analysis of upregulated Hallmark gene sets (B) and p53 and Atf4 target genes (C) in the transcriptome of both Elp3 HscKO LT-HSCs and MyPs compared with their Elp3 fl/fl counterparts (*, familywise error rate [FWER] P < 0.05; ***, P < 1.10 −3 ; ****, P < 1.10 −4 ). (D) Heatmap of the expression of the top 20 leading edge genes in the “Cell death” GO gene set in Elp3 fl/fl and Elp3 HscKO LT-HSCs and MyPs. ( E and F ) Intracellular flow cytometric (ICFC) profile of p53 (E) and phospho-Ser 15 p53 (F) and signal intensity in Elp3 fl/fl and Elp3 HscKO HSCs and MyPs (two way-ANOVA, n = 10/group, data pooled from three experiments). (G) Heatmap of the expression of the top 20 leading edge Atf4 target genes in Elp3 fl/fl and Elp3 HscKO LT-HSCs and MyPs. (H) Intracellular flow cytometric profile of Atf4 expression and comparison of the percentages of Atf4 high cells (left) and Atf4 signal intensity (right) in Elp3 fl/fl and Elp3 HscKO HSCs and MyPs (ns, not significant; ****, P < 1.10 −4 in a two-way ANOVA, n = 8–9/group, data pooled from two experiments). FDR, false discovery rate. ns, not significant; ****, P < 1.10 −4 .

Article Snippet: ELP3 rabbit mAb (clone D5H12, cat#5728, RRID:AB_11178379), β-actin mouse mAb (clone 8H10D10, cat#3700, RRID:AB_2242334), anti-mouse IgG, HRP-linked antibody (cat#7076, RRID:AB_330924), anti-rabbit IgG, HRP-linked antibody (cat#7074, RRID:AB_2099233), ATF4 rabbit mAb (clone D4B8, cat#11815S, RRID:AB_2616025), phospho-eIF2α (Ser51) rabbit mAb (clone D9G8, cat#3398S, RRID:AB_2096481), eIF2α rabbit mAb (clone D7D3, cat#5324S, RRID:AB_10692650), p53 Alexa Fluor 647 mouse mAb (clone 1C12, cat#2533, RRID:AB_2256294), phospho-p53 (Ser15) PE mouse mAB (clone 16G8, cat#8514, RRID:AB_11178384), 4E-BP1 PE rabbit mAb (clone 53H11, cat#34470, RRID:AB_2799054), phospho-4E-BP1 (Thr37/46) Alexa Fluor 488 rabbit mAb (clone 236B4, cat#2846, RRID:AB_2292749), and c-Myc Alexa Fluor 488 rabbit mAb (clone D84C12, Cat#12855, RRID:AB_2798045) were purchased from Cell Signaling Technology.

Techniques: Expressing, Comparison

Journal: The Journal of Experimental Medicine

Article Title: Loss of tRNA-modifying enzyme Elp3 activates a p53-dependent antitumor checkpoint in hematopoiesis

doi: 10.1084/jem.20200662

Figure Lengend Snippet: Deficiency in Elongator activates an amino acid deprivation-like response coordinated by p53 and Atf4 in hematopoietic progenitors. (A) Flow cytometric profiles and normalized signal intensity of dichlorodihydrofluorescein diacetate (DCFDA; left) and Mitosox (right) staining in Elp3 fl/fl and Elp3 HscKO LT-HSCs and MyPs (two-way ANOVA, n = 9–18/group, data pooled from three experiments). (B and C) GSEAR analysis of the enrichment in amino acid deprivation response genes (B; ) and heatmaps of the expression of the respective top 20 leading edge genes in the amino acid deprivation response gene set (C) in the transcriptome of Elp3 HscKO LT-HSCs (left) and MyPs (right). (D and E) Intracellular flow cytometric (ICFC) profiles and signal intensity of total 4EBP1, phospho-Thr 37/46 4EBP1 (D) and c-Myc (E) in Elp3 fl/fl and Elp3 HscKO HSCs and MyPs (two-way ANOVA, n = 10/group, data pooled from three experiments; staining controls: WT BM progenitors treated for 4 h with 100 nM rapamycin or cultured overnight in the absence of growth factors [GF]). (F) Atf4 mRNA expression in Elp3 fl/fl and Elp3 HscKO MyPs (two way-ANOVA, n = 6/group; data from and ). FMO, fluorescence minus one; NES, normalized enrichment score. ns, not significant; **, P < 0.01; ***, P < 0.001; ****, P < 1.10 −4 .

Article Snippet: ELP3 rabbit mAb (clone D5H12, cat#5728, RRID:AB_11178379), β-actin mouse mAb (clone 8H10D10, cat#3700, RRID:AB_2242334), anti-mouse IgG, HRP-linked antibody (cat#7076, RRID:AB_330924), anti-rabbit IgG, HRP-linked antibody (cat#7074, RRID:AB_2099233), ATF4 rabbit mAb (clone D4B8, cat#11815S, RRID:AB_2616025), phospho-eIF2α (Ser51) rabbit mAb (clone D9G8, cat#3398S, RRID:AB_2096481), eIF2α rabbit mAb (clone D7D3, cat#5324S, RRID:AB_10692650), p53 Alexa Fluor 647 mouse mAb (clone 1C12, cat#2533, RRID:AB_2256294), phospho-p53 (Ser15) PE mouse mAB (clone 16G8, cat#8514, RRID:AB_11178384), 4E-BP1 PE rabbit mAb (clone 53H11, cat#34470, RRID:AB_2799054), phospho-4E-BP1 (Thr37/46) Alexa Fluor 488 rabbit mAb (clone 236B4, cat#2846, RRID:AB_2292749), and c-Myc Alexa Fluor 488 rabbit mAb (clone D84C12, Cat#12855, RRID:AB_2798045) were purchased from Cell Signaling Technology.

Techniques: Staining, Expressing, Cell Culture, Fluorescence

Journal: The Journal of Experimental Medicine

Article Title: Loss of tRNA-modifying enzyme Elp3 activates a p53-dependent antitumor checkpoint in hematopoiesis

doi: 10.1084/jem.20200662

Figure Lengend Snippet: Deletion of p53 rescues Elongator-deficient hematopoiesis. (A) Hemocytometric comparison of RBCs, leukocytes, and platelets in Elp3 fl/fl , Tr p53 HscKO , Elp3 HscKO , and Elp3/Trp53 HscDKO mice (ANOVA, n = 12–14/group, data pooled from three experiments). (B–D) Total BM (B), MyP (C), and LT-HSC (D) cell counts in the femur of Elp3 fl/fl , Tr p53 HscKO , Elp3 HscKO , and Elp3/ Tr p53 HscDKO mice (ANOVA, n = 12–14/group, data pooled from three experiments). (E–G) Experimental outline (E), blood polymorphonuclear (PMN) concentrations (F), and total BM and MyPs counts (G) in 5-FU and excipient-treated Elp3 fl/fl , Tr p53 HscKO , Elp3 HscKO , and Elp3/Trp53 HscDKO mice (two-sided pairwise t tests with Bonferroni correction, n = 6/group, pool of two experiments). (H) Depiction of Elongator and CTU1/2 catalyzed tRNA modifications. (I) Flow cytometric profile of L-homopropargylglycine (L-HPG) incorporation (left) and normalized signal intensity (right) in shCTRL, shELP3, and shCTU2 TF-1 cells (two-way ANOVA, n = 7/group, data pooled from four experiments). (J) p53, ELP3, and CTU2 protein expression assessed by nanocapillary immunoassay with normalization module (left, representative pseudoblots; right, quantification of p53 protein expression) in shCTRL, shELP3, and shCTU2 TF-1 cells (two-way ANOVA, n = 13/group, data pooled from six experiments). (K) Intracellular flow cytometric profile (upper) and normalized intensities of nuclear Atf4 expression (lower) in Elp3 fl/fl , Elp3 HscKO , and Atf4 HscTG HSCs and MyPs (two-way ANOVA, n = 5/group, data pooled from two experiments). (L–N) Experimental outline (L), blood PMN concentrations (M), and total BM and MyPs counts (N) in 5-FU and excipient-treated Elp3 fl/fl , Elp3 HscKO and Atf4 HscTG , and Atf4 HscTG Trp53 fl/+ Vav1-iCre T/+ mice (pairwise t tests with Bonferroni correction, n = 5–7/group, pool of two experiments). ns, not significant; *, P < 0.05; **, P < 0.001; ***, P < 0.001; ****, P < 1.10 −4 .

Article Snippet: ELP3 rabbit mAb (clone D5H12, cat#5728, RRID:AB_11178379), β-actin mouse mAb (clone 8H10D10, cat#3700, RRID:AB_2242334), anti-mouse IgG, HRP-linked antibody (cat#7076, RRID:AB_330924), anti-rabbit IgG, HRP-linked antibody (cat#7074, RRID:AB_2099233), ATF4 rabbit mAb (clone D4B8, cat#11815S, RRID:AB_2616025), phospho-eIF2α (Ser51) rabbit mAb (clone D9G8, cat#3398S, RRID:AB_2096481), eIF2α rabbit mAb (clone D7D3, cat#5324S, RRID:AB_10692650), p53 Alexa Fluor 647 mouse mAb (clone 1C12, cat#2533, RRID:AB_2256294), phospho-p53 (Ser15) PE mouse mAB (clone 16G8, cat#8514, RRID:AB_11178384), 4E-BP1 PE rabbit mAb (clone 53H11, cat#34470, RRID:AB_2799054), phospho-4E-BP1 (Thr37/46) Alexa Fluor 488 rabbit mAb (clone 236B4, cat#2846, RRID:AB_2292749), and c-Myc Alexa Fluor 488 rabbit mAb (clone D84C12, Cat#12855, RRID:AB_2798045) were purchased from Cell Signaling Technology.

Techniques: Comparison, Expressing

Journal: The Journal of Experimental Medicine

Article Title: Loss of tRNA-modifying enzyme Elp3 activates a p53-dependent antitumor checkpoint in hematopoiesis

doi: 10.1084/jem.20200662

Figure Lengend Snippet: Deletion of p53 rescues Elongator-deficient hematopoiesis. (A) Principal component analysis on the transcriptome of Elp3 fl/fl , Tr p53 HscKO , Elp3 HscKO , and Elp3/Trp53 HscDKO MyPs. (B) Correlation between changes in mRNA expression in Elp3 HscKO vs . Elp3 fl/fl MyPs and in Elp3/Trp53 HscDKO vs . Trp53 HscKO MyPs. (C) GSEAR analysis of Atf4 target genes in genes coregulated in both Elp3 HscKO and Elp3/Trp53 HscDKO MyPs compared with their Elp3 fl/fl or Tr p53 HscKO counterparts. (D) Heatmap of the expression of the top 20 leading edge Atf4 target genes in Elp3 fl/fl , Tr p53 HscKO , Elp3 HscKO , and Elp3/Trp53 HscDKO MyPs. (E and F) Intracellular flow cytometric (ICFC) profile of Atf4 (E) and phospho-Ser51 eIF2α signal (F) in Elp3 fl/fl , Tr p53 HscKO , Elp3 HscKO , and Elp3/Trp53 HscDKO HSCs and MyPs (ns, not significant; ***, P < 0.001; ****, P < 1.10 −4 in a two-way ANOVA followed by Tukey HSD tests, n = 5–8/group, data pooled from two experiments). FDR, false discovery rate; NES, normalized enrichment score.

Article Snippet: ELP3 rabbit mAb (clone D5H12, cat#5728, RRID:AB_11178379), β-actin mouse mAb (clone 8H10D10, cat#3700, RRID:AB_2242334), anti-mouse IgG, HRP-linked antibody (cat#7076, RRID:AB_330924), anti-rabbit IgG, HRP-linked antibody (cat#7074, RRID:AB_2099233), ATF4 rabbit mAb (clone D4B8, cat#11815S, RRID:AB_2616025), phospho-eIF2α (Ser51) rabbit mAb (clone D9G8, cat#3398S, RRID:AB_2096481), eIF2α rabbit mAb (clone D7D3, cat#5324S, RRID:AB_10692650), p53 Alexa Fluor 647 mouse mAb (clone 1C12, cat#2533, RRID:AB_2256294), phospho-p53 (Ser15) PE mouse mAB (clone 16G8, cat#8514, RRID:AB_11178384), 4E-BP1 PE rabbit mAb (clone 53H11, cat#34470, RRID:AB_2799054), phospho-4E-BP1 (Thr37/46) Alexa Fluor 488 rabbit mAb (clone 236B4, cat#2846, RRID:AB_2292749), and c-Myc Alexa Fluor 488 rabbit mAb (clone D84C12, Cat#12855, RRID:AB_2798045) were purchased from Cell Signaling Technology.

Techniques: Expressing

Journal: The Journal of Experimental Medicine

Article Title: Loss of tRNA-modifying enzyme Elp3 activates a p53-dependent antitumor checkpoint in hematopoiesis

doi: 10.1084/jem.20200662

Figure Lengend Snippet: p53 is activated as an antitumor checkpoint in Elongator-deficient hematopoietic progenitors . (A) Survival curve of Elp3 fl/fl , Tr p53 HscKO , Elp3 HscKO , and Elp3/Trp53 HscDKO mice ( n = 20/group, Mantel-Cox test). (B) Survival curve of Elp3 fl/fl and Elp3 HscKO mice ( n = 18/group, Mantel-Cox test; cohort independent of that in ). (C) Representative sections of tumors of Elp3 HscKO mice (upper panel scale bars, 100 µm; lower panels, 20 µm). ( D) Types and prevalence of Trp53 exon mutations in tumors of Elp3 HscKO mice (left panel, n = 11 thymus or spleen tumors from individual mice) and position and predicted consequences of said mutations on the p53 protein (right panel: * and **, mutations cooccurring in two individual tumors; spade symbol, deletion found in tumors of two individuals; bold, predicted loss-of-function mutations). (E) Model of the consequences of Elongator inactivation on HSCs and downstream progenitors. Loss of U34 modifications activates p53 and Atf4 in what resembles a misguided amino acid deprivation response. Possibly owing to their higher metabolic activity, MyPs overactivate Atf4 and a proapoptotic gene expression program, whereas HSCs are less impacted. Mutation of the p53 checkpoint favors tumorigenesis. SNV, single nucleotide variant; H, helix. **, P < 0.01; ****, P < 1.10 −4 .

Article Snippet: ELP3 rabbit mAb (clone D5H12, cat#5728, RRID:AB_11178379), β-actin mouse mAb (clone 8H10D10, cat#3700, RRID:AB_2242334), anti-mouse IgG, HRP-linked antibody (cat#7076, RRID:AB_330924), anti-rabbit IgG, HRP-linked antibody (cat#7074, RRID:AB_2099233), ATF4 rabbit mAb (clone D4B8, cat#11815S, RRID:AB_2616025), phospho-eIF2α (Ser51) rabbit mAb (clone D9G8, cat#3398S, RRID:AB_2096481), eIF2α rabbit mAb (clone D7D3, cat#5324S, RRID:AB_10692650), p53 Alexa Fluor 647 mouse mAb (clone 1C12, cat#2533, RRID:AB_2256294), phospho-p53 (Ser15) PE mouse mAB (clone 16G8, cat#8514, RRID:AB_11178384), 4E-BP1 PE rabbit mAb (clone 53H11, cat#34470, RRID:AB_2799054), phospho-4E-BP1 (Thr37/46) Alexa Fluor 488 rabbit mAb (clone 236B4, cat#2846, RRID:AB_2292749), and c-Myc Alexa Fluor 488 rabbit mAb (clone D84C12, Cat#12855, RRID:AB_2798045) were purchased from Cell Signaling Technology.

Techniques: Activity Assay, Gene Expression, Mutagenesis, Variant Assay

Journal: The Journal of Experimental Medicine

Article Title: Loss of tRNA-modifying enzyme Elp3 activates a p53-dependent antitumor checkpoint in hematopoiesis

doi: 10.1084/jem.20200662

Figure Lengend Snippet: p53 is activated as an antitumor checkpoint in Elongator-deficient hematopoietic progenitors. (A and B) Tumor-bearing organs in Tr p53 HscKO and Elp3/Trp53 HscDKO mice reaching ethical end points (A) and representative pictures of such tumors (B; scale bars, 1 cm). (C) Incidence of tumors and tumor-bearing organs in Elp3 HscKO mice reaching ethical end points.

Article Snippet: ELP3 rabbit mAb (clone D5H12, cat#5728, RRID:AB_11178379), β-actin mouse mAb (clone 8H10D10, cat#3700, RRID:AB_2242334), anti-mouse IgG, HRP-linked antibody (cat#7076, RRID:AB_330924), anti-rabbit IgG, HRP-linked antibody (cat#7074, RRID:AB_2099233), ATF4 rabbit mAb (clone D4B8, cat#11815S, RRID:AB_2616025), phospho-eIF2α (Ser51) rabbit mAb (clone D9G8, cat#3398S, RRID:AB_2096481), eIF2α rabbit mAb (clone D7D3, cat#5324S, RRID:AB_10692650), p53 Alexa Fluor 647 mouse mAb (clone 1C12, cat#2533, RRID:AB_2256294), phospho-p53 (Ser15) PE mouse mAB (clone 16G8, cat#8514, RRID:AB_11178384), 4E-BP1 PE rabbit mAb (clone 53H11, cat#34470, RRID:AB_2799054), phospho-4E-BP1 (Thr37/46) Alexa Fluor 488 rabbit mAb (clone 236B4, cat#2846, RRID:AB_2292749), and c-Myc Alexa Fluor 488 rabbit mAb (clone D84C12, Cat#12855, RRID:AB_2798045) were purchased from Cell Signaling Technology.

Techniques: