Journal: Journal of translational medicine

Article Title: Establishment of a prognostic model based on ER stress-related cell death genes and proposing a novel combination therapy in acute myeloid leukemia.

doi: 10.1186/s12967-025-06615-y

Figure Lengend Snippet: Fig. 9 DDIT4 promotes AML cell proliferation in vitro. A Knockdown of DDIT4 in U937 and HL60 cell lines. B‒H Analysis of cell viability (B), cell counting (C), colony formation (D‒E), cell apoptosis (F), the cell cycle (G), and migration (H) in control and DDIT4-knockdown cells. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The primary antibodies used for the western blotting assays were rabbit antibodies against DDIT4 (1:1000, 10638-1- AP; Proteintech, Wuhan, Beijing) and mouse antibodies against beta-actin (1:1000, AC004; ABclonal Technology, Wuhan, China).

Techniques: In Vitro, Knockdown, Cell Counting, Migration, Control

Journal: Respiratory research

Article Title: REDD1 mediates HDM-induced nuclear-cytoplasmic translocation and release of IL-33 in airway epithelial cells by downregulating Nrf2.

doi: 10.1186/s12931-025-03119-7

Figure Lengend Snippet: Fig. 1 REDD1 expression is increased in asthma patients. REDD1 is significantly upregulated in respiratory-related diseases, positively correlates with inflammation, and negatively correlates with Nrf2. (A) Expression levels of REDD1 in sputum samples from asthma and COPD patients; (B) Correlation heatmap of REDD1 with Nrf2 and inflammatory factors in sputum samples from COPD patients; (C) Correlation heatmap of REDD1 with Nrf2 and inflam matory factors in sputum samples from asthma patients; (D) Expression levels of REDD1 in non-severe and severe asthma patients; (E) Representative immunohistochemical staining of REDD1 in the bronchial mucosal of patients with asthma as well as healthy controls. Scale bar, 100 μm

Article Snippet: The primary antibodies used in the experiments were as follows: REDD1 (1:1000; Abclonal, China), β-actin (1:10000; Wuhan Sanying, Wuhan, China), Nrf2 (1:1000; Abcam, Cambridge, UK), IL-33 (1:1000; Abclonal, China), and HO-1 (1:1000; Abclonal, China).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Respiratory research

Article Title: REDD1 mediates HDM-induced nuclear-cytoplasmic translocation and release of IL-33 in airway epithelial cells by downregulating Nrf2.

doi: 10.1186/s12931-025-03119-7

Figure Lengend Snippet: Fig. 3 HDM induces IL-33 secretion and increases expression of REDD1 and Nrf2 in 16HBE cells. (A) Detection of REDD1 and Nrf2 protein levels by Western blot in 16HBE cells stimulated with HDM (10 µg/mL) for 0 to 24 h; (B) Detection of IL-33 protein levels in the supernatant by western blot after stimulation of 16HBE cells with HDM (10 µg/mL) for 0 to 24 h. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The primary antibodies used in the experiments were as follows: REDD1 (1:1000; Abclonal, China), β-actin (1:10000; Wuhan Sanying, Wuhan, China), Nrf2 (1:1000; Abcam, Cambridge, UK), IL-33 (1:1000; Abclonal, China), and HO-1 (1:1000; Abclonal, China).

Techniques: Expressing, Western Blot

Journal: Respiratory research

Article Title: REDD1 mediates HDM-induced nuclear-cytoplasmic translocation and release of IL-33 in airway epithelial cells by downregulating Nrf2.

doi: 10.1186/s12931-025-03119-7

Figure Lengend Snippet: Fig. 4 Knocking down REDD1 alleviates HDM-induced IL-33 release and inflammatory responses in 16HBE cells. (A-B) Expression of REDD1 in 16HBE cells transfected with control, si-NC, si-REDD1-1, si-REDD1-2, or si-REDD1-3, detected by Western blot. (C) The effect of reduced REDD1 expression on IL-33 protein levels in the cell supernatant, determined by Western blot. (D) The effect of reduced REDD1 expression on TNF-α, IL-6, and IL-1β mRNA levels, quantified by qPCR. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The primary antibodies used in the experiments were as follows: REDD1 (1:1000; Abclonal, China), β-actin (1:10000; Wuhan Sanying, Wuhan, China), Nrf2 (1:1000; Abcam, Cambridge, UK), IL-33 (1:1000; Abclonal, China), and HO-1 (1:1000; Abclonal, China).

Techniques: Expressing, Transfection, Control, Western Blot

Journal: Respiratory research

Article Title: REDD1 mediates HDM-induced nuclear-cytoplasmic translocation and release of IL-33 in airway epithelial cells by downregulating Nrf2.

doi: 10.1186/s12931-025-03119-7

Figure Lengend Snippet: Fig. 5 Knocking down REDD1 enhances Nrf2 signaling in HDM-treated 16HBE cells. The effect of 24-hour incubation with HDM (10 µg/mL) on Nrf2 and HO-1 protein levels in 16HBE cells with reduced REDD1 expression, determined by Western blot.*p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The primary antibodies used in the experiments were as follows: REDD1 (1:1000; Abclonal, China), β-actin (1:10000; Wuhan Sanying, Wuhan, China), Nrf2 (1:1000; Abcam, Cambridge, UK), IL-33 (1:1000; Abclonal, China), and HO-1 (1:1000; Abclonal, China).

Techniques: Incubation, Expressing, Western Blot

Journal: Respiratory research

Article Title: REDD1 mediates HDM-induced nuclear-cytoplasmic translocation and release of IL-33 in airway epithelial cells by downregulating Nrf2.

doi: 10.1186/s12931-025-03119-7

Figure Lengend Snippet: Fig. 7 Knocking down REDD1 regulates IL-33 release through activating Nrf2. (A-B) After knockdown of REDD1 with small molecule inhibitors, cells were pretreated with ML385 followed by stimulation with HDM, and the expression of IL-33 in the supernatant was detected by Western blot.*p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The primary antibodies used in the experiments were as follows: REDD1 (1:1000; Abclonal, China), β-actin (1:10000; Wuhan Sanying, Wuhan, China), Nrf2 (1:1000; Abcam, Cambridge, UK), IL-33 (1:1000; Abclonal, China), and HO-1 (1:1000; Abclonal, China).

Techniques: Knockdown, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: DDIT4/mTOR signaling pathway mediates cantharidin-induced hepatotoxicity and cellular damage

doi: 10.3389/fphar.2024.1480512

Figure Lengend Snippet: Effects of cantharidin on mRNA and protein expression of DDIT4 in mouse liver. (A) western blot gel electrophoresis of DDIT4, with β-actin as the internal reference; (B) Immunohistochemical map of DDIT4 protein, the brown area indicated by the arrow represents the positive rate; (C) Quantitative protein map by western blot; (D) Immunohistochemical quantitative map; (E) qRT-PCR diagram of DDIT4 gene, with β-actin as the internal reference gene.

Article Snippet: The tissue protein samples were subjected to SDS-PAGE gel electrophoresis and membrane transfer, before sealing with 5% skimmed milk powder for 2 h. The following antibodies were dripped: DDIT4 (1:1000, A14135, Abclonal, China), Beclin-1 (1:1000, A7353, Abclonal, China), LC3 (1:1000, AF5402, Affinity, USA), mToR (1:1000, AF6308, Affinity, USA), p-mToR (1:1000, AF3308, Affinity, USA), Bax (1:1000, AF0120, Affinity, USA A), Bcl-2 (1:1000, AF6139, Affinity, USA) β-Actin (1:1000, BM0627, Bosterbio, China), and GAPDH (1:1000, AB-P-R 001, Goodhere, China), where β-Actin and GAPDH were used as internal references.

Techniques: Expressing, Western Blot, Nucleic Acid Electrophoresis, Immunohistochemical staining, Quantitative RT-PCR

Journal: Frontiers in Pharmacology

Article Title: DDIT4/mTOR signaling pathway mediates cantharidin-induced hepatotoxicity and cellular damage

doi: 10.3389/fphar.2024.1480512

Figure Lengend Snippet: Effects of CTD on DDIT4 expression. (A–C) : The mRNA relative expression level of the DDIT4 gene in L-O2 cells after 12, 24, and 36 h of CTD exposure was detected by qRT-PCR, and GAPDH was the internal reference gene. (D, E) : WB detected the relative expression level of DDIT4 protein in L-O2 cells after 24 h exposure to CTD.

Article Snippet: The tissue protein samples were subjected to SDS-PAGE gel electrophoresis and membrane transfer, before sealing with 5% skimmed milk powder for 2 h. The following antibodies were dripped: DDIT4 (1:1000, A14135, Abclonal, China), Beclin-1 (1:1000, A7353, Abclonal, China), LC3 (1:1000, AF5402, Affinity, USA), mToR (1:1000, AF6308, Affinity, USA), p-mToR (1:1000, AF3308, Affinity, USA), Bax (1:1000, AF0120, Affinity, USA A), Bcl-2 (1:1000, AF6139, Affinity, USA) β-Actin (1:1000, BM0627, Bosterbio, China), and GAPDH (1:1000, AB-P-R 001, Goodhere, China), where β-Actin and GAPDH were used as internal references.

Techniques: Expressing, Quantitative RT-PCR

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