Journal: Cell Reports

Article Title: Oxidative Stress Triggers Selective tRNA Retrograde Transport in Human Cells during the Integrated Stress Response

doi: 10.1016/j.celrep.2019.02.077

Figure Lengend Snippet:

Article Snippet: Rabbit Antibody against REDD1/DDIT4 , Novus Biologicals , Cat# NBP1-77321SS; RRID: AB_11036185.

Techniques: Recombinant, Cell Isolation, Reverse Transcription, SYBR Green Assay, Isolation, Sequencing, Software

Journal: The Journal of experimental biology

Article Title: Different fuel regulation in two types of myofiber results in different antioxidant strategies in Daurian ground squirrels ( Spermophilus dauricus ) during hibernation.

doi: 10.1242/jeb.231639

Figure Lengend Snippet: Fig. 1. Expression of REDD1 in SOL and EDL muscles of Daurian ground squirrels during hibernation. Protein expression levels were visualized at four sampling points: pre-hibernation (PRE), hibernation (HIB), interbout arousal (IBA) and post-hibernation (POST). See Materials and Methods for more extensive definitions. Representative immunoblots and Stain-Free total protein loading controls are shown below for selected pairs of sampling points and muscles that are labeled on the top of the blot. Data were analyzed using an ANOVA with a post hoc Tukey’s test (P<0.05); values that are not statistically different from each other share the same letter notation.

Article Snippet: Membranes were then blocked with 4% Mowiol® PVA-203 (Aladdin, China) in Tris-buffered saline (TBS) for 10 min (Gholap et al., 2005; Rodda and Yamazaki, 1994), and incubated with REDD1 (Proteintech, 10638-1-AP), atrogin-1 (Proteintech, 12866-1-AP), PGC-1α (Novus, NBP1-04676SS), FOXO1 (C29H4) rabbit mAb (CST#2880), phospho-FOXO1 (Ser256) antibody (CST#9461), FOXO4 (CST#9472) and FOXO4 (phospho-Thr451) (Signalway antibody, 12053) in 0.1% TBST (TBS with 0.1% Tween 20) containing 2% polyvinylpyrrolidone (PVP-40; Amresco, USA, 0507-500G) at 4°C overnight.

Techniques: Expressing, Muscles, Sampling, Western Blot, Staining, Labeling

Journal: Discover oncology

Article Title: Increased nuclear expression of DNA damage inducible transcript 4 can serve as a potential prognostic biomarker in patients with gliomas: a study based on data mining and experimental tools.

doi: 10.1007/s12672-025-01865-0

Figure Lengend Snippet: Fig. 1 Genetic alteration analysis of DDIT4 in glioma patients using cBioPortal. The analysis indicates that the DDIT4 gene is altered in less than 1% of patients. Alterations include copy number variations (CNV) such as deep deletions and somatic mutations, with no significant representation of other genetic changes

Article Snippet: Subsequently, the sections were incubated with a primary antibody specific for DDIT4 (1:80 dilution, Biorbyt, Cambridge, MA, UK) for an overnight at 4◦C.

Techniques:

Journal: Discover oncology

Article Title: Increased nuclear expression of DNA damage inducible transcript 4 can serve as a potential prognostic biomarker in patients with gliomas: a study based on data mining and experimental tools.

doi: 10.1007/s12672-025-01865-0

Figure Lengend Snippet: Fig. 4 GeneMANIA network analysis for DDIT4 and associated genes. The analysis identifies gene sets enriched in the DDIT4 network, rep- resented by various edge colors indicating different types of interactions: Physical Interactions, Co-expression, Predicted, Co-localization, Genetic Interactions, Pathway, and Shared protein domains. Node colors correspond to the biological functions of the enriched gene sets, such as TOR signaling and response to oxygen levels

Article Snippet: Subsequently, the sections were incubated with a primary antibody specific for DDIT4 (1:80 dilution, Biorbyt, Cambridge, MA, UK) for an overnight at 4◦C.

Techniques: Expressing

Journal: Discover oncology

Article Title: Increased nuclear expression of DNA damage inducible transcript 4 can serve as a potential prognostic biomarker in patients with gliomas: a study based on data mining and experimental tools.

doi: 10.1007/s12672-025-01865-0

Figure Lengend Snippet: Fig. 3 Prognostic analysis of DDIT4 mRNA expression in glioma patients using the GEPIA2 tool. High DDIT4 expression is significantly asso- ciated with worse prognosis in gliomas, including glioblastoma multiforme (GBM) and low-grade gliomas (LGG). A Kaplan–Meier curve for overall survival (OS) shows that patients with high DDIT4 expression have reduced survival compared to those with low expression. B Kaplan–Meier curve for recurrence-free survival (RFS) demonstrates a similar trend, with high DDIT4 expression correlating with shorter recurrence-free periods. Statistical significance is indicated with log-rank P-values

Article Snippet: Subsequently, the sections were incubated with a primary antibody specific for DDIT4 (1:80 dilution, Biorbyt, Cambridge, MA, UK) for an overnight at 4◦C.

Techniques: Expressing

Journal: Discover oncology

Article Title: Increased nuclear expression of DNA damage inducible transcript 4 can serve as a potential prognostic biomarker in patients with gliomas: a study based on data mining and experimental tools.

doi: 10.1007/s12672-025-01865-0

Figure Lengend Snippet: Fig. 5 Immunohistochemical (IHC) analysis of DDIT4 protein expression in glial tumors and controls. A, A-1, A-2 Positive nuclear expression of DDIT4 in glial tumor tissues at different magnifications (100×, 200×, and 400×). B, B-1, B-2 Positive cytoplasmic expression of DDIT4 in glial tumor tissues at corresponding magnifications. C IHC staining of normal glial tissue. D Human normal kidney tissue as a positive con- trol. E Human normal kidney tissue as a negative control. F Isotype control for validation. Magnifications are indicated for each panel

Article Snippet: Subsequently, the sections were incubated with a primary antibody specific for DDIT4 (1:80 dilution, Biorbyt, Cambridge, MA, UK) for an overnight at 4◦C.

Techniques: Immunohistochemical staining, Expressing, Immunohistochemistry, Negative Control, Control, Biomarker Discovery

Journal: Discover oncology

Article Title: Increased nuclear expression of DNA damage inducible transcript 4 can serve as a potential prognostic biomarker in patients with gliomas: a study based on data mining and experimental tools.

doi: 10.1007/s12672-025-01865-0

Figure Lengend Snippet: Fig. 6 Kaplan–Meier survival curves for disease-specific survival (DSS) and recurrence- free survival (RFS) based on nuclear DDIT4 protein expres- sion levels in glial tumors. A Kaplan–Meier analysis for DSS indicates that tumors with positive nuclear DDIT4 expression are associated with significantly worse survival compared to those with negative expression (Log-rank test, P = 0.013). B Kaplan–Meier analysis for RFS reveals a significant associa- tion between positive nuclear DDIT4 expression and shorter recurrence-free periods (Log- rank test, P = 0.024). Survival statistics include mean and median values with 95% con- fidence intervals, as shown in the accompanying tables

Article Snippet: Subsequently, the sections were incubated with a primary antibody specific for DDIT4 (1:80 dilution, Biorbyt, Cambridge, MA, UK) for an overnight at 4◦C.

Techniques: Expressing

Journal: Discover oncology

Article Title: Increased nuclear expression of DNA damage inducible transcript 4 can serve as a potential prognostic biomarker in patients with gliomas: a study based on data mining and experimental tools.

doi: 10.1007/s12672-025-01865-0

Figure Lengend Snippet: Fig. 7 Kaplan–Meier survival curves for disease-specific survival (DSS) (A) and recur- rence-free survival (RFS) (B) in patients with glial tumors treated with temozolomide (TMZ). The analysis indicates no significant differences in DSS or RFS between patients with positive and nega- tive nuclear DDIT4 expres- sion under TMZ treatment (Log-rank test: P = 0.129 and P = 0.299, respectively)

Article Snippet: Subsequently, the sections were incubated with a primary antibody specific for DDIT4 (1:80 dilution, Biorbyt, Cambridge, MA, UK) for an overnight at 4◦C.

Techniques:

Journal: Discover oncology

Article Title: Increased nuclear expression of DNA damage inducible transcript 4 can serve as a potential prognostic biomarker in patients with gliomas: a study based on data mining and experimental tools.

doi: 10.1007/s12672-025-01865-0

Figure Lengend Snippet: Fig. 8 Kaplan–Meier survival curves for disease-specific sur- vival (DSS) (A) and recurrence- free survival (RFS) (B) based on cytoplasmic DDIT4 protein expression levels in glial tumors. Kaplan–Meier analysis reveals no significant differ- ences in DSS or RFS between patients with positive and negative cytoplasmic DDIT4 expression (Log-rank test: P = 0.884 for DSS and P = 0.974 for RFS). Survival statistics, including mean and median values with 95% confidence intervals, are displayed in the accompanying tables

Article Snippet: Subsequently, the sections were incubated with a primary antibody specific for DDIT4 (1:80 dilution, Biorbyt, Cambridge, MA, UK) for an overnight at 4◦C.

Techniques: Expressing

Journal: Ecotoxicology and environmental safety

Article Title: DDIT4 is essential for DINP-induced autophagy of ovarian granulosa cells.

doi: 10.1016/j.ecoenv.2023.115686

Figure Lengend Snippet: Fig. 2. DINP upregulates DDIT4 expression in ovarian granulosa cells. (A) DDIT4 expression in the ovary tissue was detected by IHC after adult female mice were administered with 0–200 mg/kg DINP for 14 days. The scale bar was 100 µm. (B) The mRNA level of DDIT4 was determined by qPCR after KGN cells were exposed to 0 or 800 μM DINP for 24 h. (C, D) The protein expression of DDIT4 was detected by Western blot after KGN cells were exposed to 0–800 μM DINP for 24 h. *P < 0.05.

Article Snippet: Rabbit anti-DDIT4 polyclonal antibody (10638–1-AP) and mouse IgG (B900620) were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Expressing, Western Blot

Journal: Ecotoxicology and environmental safety

Article Title: DDIT4 is essential for DINP-induced autophagy of ovarian granulosa cells.

doi: 10.1016/j.ecoenv.2023.115686

Figure Lengend Snippet: Fig. 3. DDIT4 is essential for DINP-induced autophagy of ovarian granulosa cells. (A, B) The protein levels of LC3, Beclin 1, Atg 5, p62 and DDIT4 were determined by Western blot after KGN cells were transfected with 0, 1, 2 μg pcDNA3.1-DDIT4 plasmid for 48 h. (C) Autophagic vesicles were observed by TEM after KGN cells were transfected with 0 or 2 μg pcDNA3.1-DDIT4 for 48 h. (D, E) The expression of LC3, Beclin 1, Atg 5, p62 and DDIT4 in KGN cells was detected by Western blot after the cells were transfected with si-NC or si-DDIT4 for 24 h. The protein levels of LC3, Beclin 1, Atg 5, p62 and DDIT4 (F, G) and the DDIT4 mRNA level (H) in KGN cells were detected after the cells were exposed to 0 or 800 μM DINP in the presence or absence of si-DDIT4 for 24 h.*P < 0.05.

Article Snippet: Rabbit anti-DDIT4 polyclonal antibody (10638–1-AP) and mouse IgG (B900620) were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Western Blot, Transfection, Plasmid Preparation, Expressing

Journal: Ecotoxicology and environmental safety

Article Title: DDIT4 is essential for DINP-induced autophagy of ovarian granulosa cells.

doi: 10.1016/j.ecoenv.2023.115686

Figure Lengend Snippet: Fig. 4. ATF4 promotes gene transcription of DDIT4. (A) Three potential ATF4 responsive elements (RE) in the DDIT4 promoter region were predicted by JASPAR. (B- D) The mRNA and protein levels of ATF4 and DDIT4 were detected by qPCR and Western blot, respectively, after KGN cells were transfected with 0, 1, 2 μg pcDNA3.1-ATF4 for 24 or 48 h. (E-G) The mRNA and protein levels of ATF4 and DDIT4 were determined by qPCR and Western blot, respectively, after KGN cells were transfected with si-NC or si-ATF4 for 24 h. (H) Luciferase activity was detected after KGN cells were transfected with pGL4.2 or pGL4.2-DDIT4 prom together with pcDNA3.1 or pcDNA3.1-ATF4 for 48 h. (I) Luciferase activity was determined after KGN cells were transfected with pGL4.2-RE1, pGL4.2-RE2 or pGL4.2-RE3, respectively, together with pcDNA3.1 or pcDNA3.1-ATF4 for 48 h. (J) Luciferase activity was determined after KGN cells were transfected with pGL4.2-RE2 or pGL4.2-mutRE2 for 48 h in the presence or absence of pcDNA3.1-ATF4. (K) ChIP-qPCR analysis was utilized to identify the binding of ATF4 to the RE2 site. *P < 0.05.

Article Snippet: Rabbit anti-DDIT4 polyclonal antibody (10638–1-AP) and mouse IgG (B900620) were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Western Blot, Transfection, Luciferase, Activity Assay, ChIP-qPCR, Binding Assay

Journal: Ecotoxicology and environmental safety

Article Title: DDIT4 is essential for DINP-induced autophagy of ovarian granulosa cells.

doi: 10.1016/j.ecoenv.2023.115686

Figure Lengend Snippet: Fig. 5. ATF4 is involved in DINP-induced upregulation of DDIT4 in ovarian granulosa cells. (A) The ATF4 expression in the ovary tissue was observed by IHC after adult female mice were administered with 0–200 mg/kg DINP for 14 d. The scale bar was 20 µm. (B-D) The ATF4 mRNA and protein level were detected by qPCR and Western blot, respectively, after KGN cells were treated with the indicated concentration of DINP for 24 h. (E-G) The expression of DDIT4 and ATF4 was detected by qPCR and Western blot, respectively, after KGN cells were exposed to 0 or 800 μM DINP for 24 h in the presence or absence of si-ATF4. *P < 0.05.

Article Snippet: Rabbit anti-DDIT4 polyclonal antibody (10638–1-AP) and mouse IgG (B900620) were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Expressing, Western Blot, Concentration Assay

Journal: Ecotoxicology and environmental safety

Article Title: DDIT4 is essential for DINP-induced autophagy of ovarian granulosa cells.

doi: 10.1016/j.ecoenv.2023.115686

Figure Lengend Snippet: Fig. 6. DINP induces autophagy of ovarian granulosa cells via ATF4/DDIT4 signals. (A, B) The protein levels of LC3, Atg 5, Beclin 1, p62 and ATF4 were determined after KGN cells were transfected with 0, 1, 2 μg pcDNA3.1-ATF4 for 48 h. (C) Autophagic vesicles were detected by TEM after KGN cells were transfected with 0 or 2 μg pcDNA3.1-ATF4 for 48 h. (D, E) The protein levels of LC3, Beclin 1, Atg 5, p62 and ATF4 were detected after KGN cells were transfected with si-NC or si-ATF4 for 24 h. (F, G) The protein levels of LC3, Beclin 1, Atg 5, p62 and ATF4 were determined after KGN cells were exposed to 0 or 800 μM DINP for 24 h in the presence or absence of si-ATF. *P < 0.05.

Article Snippet: Rabbit anti-DDIT4 polyclonal antibody (10638–1-AP) and mouse IgG (B900620) were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Transfection

Journal: PLoS Pathogens

Article Title: Influenza virus differentially activates mTORC1 and mTORC2 signaling to maximize late stage replication

doi: 10.1371/journal.ppat.1006635

Figure Lengend Snippet: ( A-C ) A549 cells were infected with WSN at MOI of 2 PFU/cell for the indicated times. Cell extracts were subjected to ( A ) western blot analysis to detect the depicted proteins and quantified as shown in or ( B ) RNA was purified for qRT-PCR to determine REDD1 mRNA levels. Mean and standard deviation are shown for qRT-PCR, n = 4 independent experiments done in triplicates. ** p <0.000004, Student's t -test. ( C ) A549 cells were transfected with siRNAs (pool of three each) targeting viral mRNAs and then infected for 7 h at MOI of 2 PFU/cell. Immunoblot analysis was performed to detect the depicted proteins, n = 3. ( D ) A549 cells were transfected with plasmids encoding the indicated virus proteins. At 48 h post-transfection, total RNA was purified and REDD1 mRNA levels were determined by qRT-PCR as in B . The bottom panel in D shows viral polymerase activity upon tranfection of the depicted viral proteins and/or minigenome as control. Minigenome mRNA was measured by qRT-PCR. In cells transfected with the complete set of plasmids that encode the viral polymerase we detect the minigenome RNA transcribed by pol I directly from the plasmid in addition to the minigenme RNA amplified by the influenza proteins, indicating protein activity. In cells transfected with the same plasmids except for PA, we only detect the minigenome RNA transcribed by pol I directly from the plasmid, and the average values was set to 1. The minigenome RNA level is higher when all plasmids were transfected ( n = 3). ( E, F ) MDCK cells were transfected with control plasmid of plasmid enconding the M2 protein. In E, RNA was purified for qRT-PCR to determine REDD1 mRNA levels as in B , n = 3, ***p<0.001. In F, cell extracts were subjected to western blot analysis to detect the depicted proteins ( n = 3). ( G ) U2OS-REDD1 cells were treated with vehicle or 1μg/ml tetracycline for 2 h prior to and during infection to induce REDD1 expression. Cells were infected at MOI of 2 PFU/cell for 6 h. Immunoblot analyses were performed to detect the depicted proteins. Total S6K serves as the loading control. The upper band in the S6K/p-S6K blots is p85 S6K, whereas the lower band is p70 S6K ( n = 3).

Article Snippet: Additional antibodies used for western blot analysis were against Rictor (Millipore 05–1471), IFITM3 (R&D Systems AF3377), MAVS (generated by Z. Chen laboratory), β-actin (Sigma A5441), REDD1 (Novus Biologicals NBP1-22966), ATG5 (Novus Biologicals NB110-53818), ATG7 (Sigma A2856), and LC3 (Novus Biologicals NB100-2220).

Techniques: Infection, Western Blot, Purification, Quantitative RT-PCR, Standard Deviation, Transfection, Virus, Activity Assay, Control, Plasmid Preparation, Amplification, Expressing

Journal: PLoS Pathogens

Article Title: Influenza virus differentially activates mTORC1 and mTORC2 signaling to maximize late stage replication

doi: 10.1371/journal.ppat.1006635

Figure Lengend Snippet: The viral protein HA and virus replication promote mTORC1 activation through PDPK1-mediated phosphorylation of AKT at T308. In addition, down-regulation of REDD1 by the viral M2 protein amplifies or support mTORC1 activation downstream of AKT. NS1 promotes AKT phosphorylation at S473 via mTORC2 and this process is known to regulate apoptosis. Differential AKT phosphorylation dictates downstream effects.

Article Snippet: Additional antibodies used for western blot analysis were against Rictor (Millipore 05–1471), IFITM3 (R&D Systems AF3377), MAVS (generated by Z. Chen laboratory), β-actin (Sigma A5441), REDD1 (Novus Biologicals NBP1-22966), ATG5 (Novus Biologicals NB110-53818), ATG7 (Sigma A2856), and LC3 (Novus Biologicals NB100-2220).

Techniques: Virus, Activation Assay, Phospho-proteomics