Journal: Scientific reports

Article Title: microRNA let-7c is essential for the anisomycin-elicited apoptosis in Jurkat T cells by linking JNK1/2 to AP-1/STAT1/STAT3 signaling.

doi: 10.1038/srep24434

Figure Lengend Snippet: Figure 1. Low dose anisomycin strongly induces the apoptosis in Jurkat T cells. Two hours after pre- incubated with or without 10 μM SP600125 (SP) or 20 μM PD98059 (PD), Jurkat T cells were treated with the indicated concentrations of anisomycin (Ani) for 6 h or 24 h or with 40 ng/ml of anisomycin for the indicated time periods. (A,B) DNA fragmentation in the treated cells was analyzed by 1% agarose gel electrophoresis. (C–E) Apoptotic proportion in the treated cells was assayed by flow cytometry using annexin-V/PI double staining. The results show the typical experiment, which has been repeated three times.

Article Snippet: After the treatments, their fragmented DNAs were extracted and purified using Apoptotic DNA Ladder Kit (Beyotime, Shanghai, China) according to the manufacturer’s instructions.

Techniques: Incubation, Agarose Gel Electrophoresis, Flow Cytometry, Double Staining

Journal: Scientific reports

Article Title: microRNA let-7c is essential for the anisomycin-elicited apoptosis in Jurkat T cells by linking JNK1/2 to AP-1/STAT1/STAT3 signaling.

doi: 10.1038/srep24434

Figure Lengend Snippet: Figure 2. JNK1/2 signaling plays the major role in the MAPK-mediated apoptosis in Jurkat T cells by anisomycin. (A–F) Jurkat T cells were treated with the increasing concentrations of anisomycin for 1 h or with 40 ng/ml of anisomycin at the indicated time. Following the treatment, the expressions of ERK1/2, P-ERK1/2 (A,B), p38, P-p38 (C,D), JNK1/2 and P-JNK1/2 (E,F) proteins were determined by Western blotting. (G,H) The cells were pretreated with 10 μM SP600125 (SP), 20 μM PD98059 (PD), 20 μM SB203580 (SB) or the increasing concentrations of SP600125 before the exposure to 40 ng/ml of anisomycin. Then, the expressions of JNK1/2 and P-JNK1/2 were examined by Western blotting. (I,J) Simultaneously, the apoptotic phenotypes of the cells treated above were observed using annexin-V/PI double staining under the flow cytometer. Data are presented as the mean ± SD of three independent experiments. *P < 0.05, ** P < 0.01 vs. the untreated control, #P < 0.05, ##P < 0.01 vs. the 40 ng/ml anisomycin group.

Article Snippet: After the treatments, their fragmented DNAs were extracted and purified using Apoptotic DNA Ladder Kit (Beyotime, Shanghai, China) according to the manufacturer’s instructions.

Techniques: Western Blot, Double Staining, Flow Cytometry, Control

Journal: Scientific reports

Article Title: microRNA let-7c is essential for the anisomycin-elicited apoptosis in Jurkat T cells by linking JNK1/2 to AP-1/STAT1/STAT3 signaling.

doi: 10.1038/srep24434

Figure Lengend Snippet: Figure 5. miRNA let-7c overexpression facilitates the anisomycin-stimulated apoptosis in Jurkat T cells. Jurkat T cells were electro-transfected with 2 μg of the let-7c overexpression vector with GFP marker (let-7c) or 2 μg of the miR-Negative Control vector (mock) for 24 h or 48 h. (A) GFP expression in Jurkat T cells. (B) The expression of miR let-7c in the transfected cells was measured by real-time qPCR. (C) The levels of phospho-c- jun, phospho-STAT1 and phospho-STAT3 were analyzed by Western blot. (D) The apoptotic rate of the treated cells was assayed by flow cytometry. The data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 vs. the untreated control, #p < 0.05, ##p < 0.01 vs. the let-7c overexpression group.

Article Snippet: After the treatments, their fragmented DNAs were extracted and purified using Apoptotic DNA Ladder Kit (Beyotime, Shanghai, China) according to the manufacturer’s instructions.

Techniques: Over Expression, Transfection, Plasmid Preparation, Marker, Negative Control, Expressing, Western Blot, Flow Cytometry, Control

Journal: Scientific reports

Article Title: microRNA let-7c is essential for the anisomycin-elicited apoptosis in Jurkat T cells by linking JNK1/2 to AP-1/STAT1/STAT3 signaling.

doi: 10.1038/srep24434

Figure Lengend Snippet: Figure 7. Role of Bim in the anisomycin-induced apoptosis in Jurkat T cells. The Jurkat T cells were transfected with 100 nM of Bim-targeting siRNA or control siRNA for 24 h to knockdown the bim gene. Then, 40 ng/ml of anisomycin was added into the cells for 24 or 48 h. (A) The Bim mRNA expression was evaluated by the real-time qRT-PCR. (B) The level of Bim protein was determined by Western blotting. (C–E) The levels of active Bak, Bax and active Bax were also measured through Western blotting. (F) The apoptotic proportion of the treated cells was analyzed by flow cytometry. (G) In situ immunofluorescence staining was performed for the changes of the active Bak and Bax in the treated cells (× 200). The data are presented as the mean ± SD of three independent experiments. *p < 0.05 and **p < 0.01 vs. the untreated control, Δp < 0.05 and ΔΔp < 0.01 vs. the anisomycin group, #p < 0.05 and ##p < 0.01 vs. the Bim siRNA group.

Article Snippet: After the treatments, their fragmented DNAs were extracted and purified using Apoptotic DNA Ladder Kit (Beyotime, Shanghai, China) according to the manufacturer’s instructions.

Techniques: Transfection, Control, Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, In Situ, Immunofluorescence, Staining

Journal: GM crops & food

Article Title: Development of transgenic wheat plants withstand salt stress via the MDAR1 gene.

doi: 10.1080/21645698.2025.2463139

Figure Lengend Snippet: Figure 2. PCR products for the (a) MDAR1 gene (520 bp) and (b) bar gene (693 bp) of the six putative transgenic plants (MD1, MD2, MD3, MD4, MD5 and MD6); M, 1Kb plus DNA ladder (Fermentas); +ve, pAb11/mdar1/bar construct; and -ve, a non-transgenic Bobwhite 56.

Article Snippet: M; New England Biolabs’ 1 kb DNA ladder containing 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, and 10 kb; (−ve) non-transgenic Bobwhite 56. (WT) subjected to salt stress (200 mm NaCl) as well as fresh water were measured to evaluate the effects of MDAR1 overexpression on ascorbic acid’s influence on H2O2 and Superoxide anion (O2•─).

Techniques: Transgenic Assay, Construct

Journal: GM crops & food

Article Title: Development of transgenic wheat plants withstand salt stress via the MDAR1 gene.

doi: 10.1080/21645698.2025.2463139

Figure Lengend Snippet: Figure 3. Southern blot investigation of the gene sequences of six MDAR1 transgenic plants (MD1 to MD6) in the T0 generation. M; New England Biolabs’ 1 kb DNA ladder containing 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, and 10 kb; (−ve) non-transgenic Bobwhite 56.

Article Snippet: M; New England Biolabs’ 1 kb DNA ladder containing 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, and 10 kb; (−ve) non-transgenic Bobwhite 56. (WT) subjected to salt stress (200 mm NaCl) as well as fresh water were measured to evaluate the effects of MDAR1 overexpression on ascorbic acid’s influence on H2O2 and Superoxide anion (O2•─).

Techniques: Southern Blot, Transgenic Assay

Journal: The Journal of biological chemistry

Article Title: Promoter characterization of the rat gene for Ca2+-binding protein regucalcin. Transcriptional regulation by signaling factors.

doi: 10.1074/jbc.274.3.1277

Figure Lengend Snippet: FIG. 1. Gel electrophoresis of PCR products of oligo-capped regucalcin mRNA. Rat liver cap site cDNA was used as a PCR tem- plate to identify the transcriptional start site of the rat regucalcin gene. The first round of PCR was performed using 1RC as the sense DNA primer complementary to r-oligo and GSAP3 as the gene-specific anti- sense primer. An aliquot of the initial PCR reaction served the template for nested PCR. The resulting cDNA extending to the cap site was amplified by nested PCR using 2RC as the nested sense DNA primer complementary to r-oligo and either GSAP1 or GSAP2 as the nested gene-specific antisense primer. The nested PCR products were electro- phoresed in 2% agarose gels and stained with ethidium bromide. The positions of GSAP1, -2, and -3 are indicated in Fig. 2. A 100-bp DNA ladder was used as the molecular size marker. Lane 1, 100-bp ladder; lane 2, nested PCR product (approximately 220 bp) using 2RC/GSAP1; lane 3, nested PCR product (approximately 250 bp) using 2RC/GSAP2.

Article Snippet: A 100-base pair DNA ladder was obtained from New England Biolabs (Beverly, MA).

Techniques: Nucleic Acid Electrophoresis, Nested PCR, Amplification, Staining, Marker