rabbit anti ddit4  (Cell Signaling Technology Inc)

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: 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