Journal: Scientific Reports

Article Title: DNA damage response induced by Etoposide promotes steroidogenesis via GADD45A in cultured adrenal cells

doi: 10.1038/s41598-018-27938-5

Figure Lengend Snippet: Inhibitory effect of GADD45A siRNA on steroidogenesis promoted by EP. H295R cells were transfected with the indicated siRNAs simultaneously with plating. After 24 h, the cells were treated with EP (0.75 μM) for 72 h, changed to the normal growth medium, and cultured for another 24 h. ( A ) Immunostaining for GADD45A and CYP21A2. H295R cells were fixed with 4% paraformaldehyde. Green staining shows the anti-GADD45A antibody, red staining shows the anti-CYP21A2 antibody, and blue staining shows DAPI (cell nuclei). Scale bars represent 100 μm. ( B ) Cortisol concentration in the medium was measured using ELISA, and corrected by the number of cells. Data are presented as the mean ± SE of 3 independent experiments. *P < 0.05 vs. control, † P < 0.05 vs. EP. ( C ) Relative mRNA expression of the indicated genes was analyzed by RT-qPCR. RNA was extracted at 12 h after changing of the medium (at 84 h after EP-treatment). Data are presented as the mean ± SE of 3 independent experiments. *P < 0.05 vs. control, † P < 0.05 vs. EP + siControl.

Article Snippet: At 1 day after plating, pcDNA3.1Zeo (+) (empty vector) or pCMV3-human GADD45A-C-FLAG vector (Sino Biological, Inc., Beijing, China) was introduced into H295R cells using Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA).

Techniques: Transfection, Cell Culture, Immunostaining, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: DNA damage response induced by Etoposide promotes steroidogenesis via GADD45A in cultured adrenal cells

doi: 10.1038/s41598-018-27938-5

Figure Lengend Snippet: Effect of GADD45A expression on steroidogenesis. H295R cells were transiently transfected with the empty vector or human GADD45A expression vector, and cultured for another 48 h. ( A ) Relative mRNA expression of the indicated genes was analyzed by qRT-PCR. Data are presented as the mean ± SE of 3 independent experiments. *P < 0.05 vs. empty. ( B ) Immunostaining for FLAG and CYP21A2. Green staining shows the anti-FLAG antibody, red staining shows the anti-CYP21A2 antibody, and blue staining shows DAPI (nuclei). Scale bars represent 100 μm. ( C ) The percentages of steroidogenic CYP21A2-positive cells in empty vector-transfected cells (Empty), total FLAG-GADD45A transfected cells (Total), FLAG-GADD45A transfected FLAG-positive [FLAG (+)] or FLAG-GADD45A transfected FLAG-negative [FLAG (−)] cells were measured. At least 200 cells from random fields were examined in each condition in a blinded manner. Data are presented as the mean ± SE of 3 independent experiments. *P < 0.05 vs. Empty, **P < 0.05 vs. Total, ***P < 0.05 vs. FLAG (−).

Article Snippet: At 1 day after plating, pcDNA3.1Zeo (+) (empty vector) or pCMV3-human GADD45A-C-FLAG vector (Sino Biological, Inc., Beijing, China) was introduced into H295R cells using Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA).

Techniques: Expressing, Transfection, Plasmid Preparation, Cell Culture, Quantitative RT-PCR, Immunostaining, Staining

Journal: Scientific Reports

Article Title: DNA damage response induced by Etoposide promotes steroidogenesis via GADD45A in cultured adrenal cells

doi: 10.1038/s41598-018-27938-5

Figure Lengend Snippet: Inhibitory effect of GADD45A siRNA on p38MAPK phosphorylation. H295R cells were transfected with the indicated siRNAs. After 24 h, the cells were treated with EP (0.75 μM) for 72 h, changed to the normal growth medium, and cultured for another 24 h. ( A ) Western blot analysis of phosphorylated p38MAPK (p-p38MAPK) and total p38MAPK (p38MAPK). Cropped images are shown. Uncropped images are shown in Supplementary Fig. . ( B ) The level of phosphorylated p38MAPK was normalized by p38MAPK expression. Quantification of the expression level was performed by ImageJ software. Data are presented as the mean ± SE of 3 independent experiments. *P < 0.05 vs. control. † P < 0.05 vs. EP + siControl.

Article Snippet: At 1 day after plating, pcDNA3.1Zeo (+) (empty vector) or pCMV3-human GADD45A-C-FLAG vector (Sino Biological, Inc., Beijing, China) was introduced into H295R cells using Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA).

Techniques: Transfection, Cell Culture, Western Blot, Expressing, Software

Journal: Frontiers in Oncology

Article Title: Radiomics based on MRI in predicting lymphovascular space invasion of cervical cancer: a meta-analysis

doi: 10.3389/fonc.2024.1425078

Figure Lengend Snippet: The basic information of literatures.

Article Snippet: Hua QW, 2022 ( ) , Retro , Siemens, 3.0 T , T1WI-CE-axis、T2WI-sag , 167 , 111 , 56 , Manually , 3D , SVM , NA.

Techniques: Sequencing, Biomarker Discovery

Journal: The Journal of Cell Biology

Article Title: Structural and functional analysis of Nup133 domains reveals modular building blocks of the nuclear pore complex

doi: 10.1083/jcb.200408109

Figure Lengend Snippet: Nucleoporins containing β-propeller domains

Article Snippet: Full-length and mutant Nup133 proteins (amino acids 1–1156, 1–514, and 497–1156) were in vitro transcribed and translated in the presence of [ 35 S]methionine by a coupled reticulocyte lysate transcription/translation system (TNT T7; Promega).

Techniques:

Journal: The Journal of Cell Biology

Article Title: Structural and functional analysis of Nup133 domains reveals modular building blocks of the nuclear pore complex

doi: 10.1083/jcb.200408109

Figure Lengend Snippet: Domain structure of Nup133. (a) Two domains were predicted, with the NTD having helical and sheet content, and the CTD being all helical. The construct used for crystallization is boxed. (b) In vitro binding of hNup133 domains to recombinant GST-hNup107. Top and bottom panels show the bound and unbound fractions of various [ 35 S]methionine-labeled Nup133 translation products incubated with recombinant GST-Nup107 immobilized on affinity resin. (c) Localization of EGFP-tagged hNup133 domains in HeLa cells. The CTD shows punctate rim staining that overlaps with the mAb414 signal. The NTD shows diffuse EGFP-signal, with a weak concentration at the nuclear rim.

Article Snippet: Full-length and mutant Nup133 proteins (amino acids 1–1156, 1–514, and 497–1156) were in vitro transcribed and translated in the presence of [ 35 S]methionine by a coupled reticulocyte lysate transcription/translation system (TNT T7; Promega).

Techniques: Construct, Crystallization Assay, In Vitro, Binding Assay, Recombinant, Labeling, Incubation, Staining, Concentration Assay

Journal: The Journal of Cell Biology

Article Title: Structural and functional analysis of Nup133 domains reveals modular building blocks of the nuclear pore complex

doi: 10.1083/jcb.200408109

Figure Lengend Snippet: Structural alignment of the seven blades in the Nup133 β-propeller. (a and b) Superposition of 21 C α positions from each blade calculated by the program MultiProt ( http://bioinfo3d.cs.tau.ac.il/MultiProt/ ). Gray spheres mark the aligned C α atoms from blade 3. Strand A lines the inner channel of the propeller and is roughly parallel to the pseudo-sevenfold axis. Blade 2 (orange), blade 5 (cyan), and blade 7 (purple) contain insertions. (c) Structure-based sequence alignment of Nup133 NTD blades. Conserved hydrophobic positions are shaded yellow. Clathrin NTD propeller blades are shown for comparison. The sequence is shown in Joy formatting ( http://www-cryst.bioc.cam.ac.uk/~joy/ ). β-Strands are shown in blue, 3 10 helices are shown in maroon, capitalized residues are solvent inaccessible, bold indicates a sidechain-backbone amide H-bond, underline indicates sidechain-backbone carbonyl H-bond, and italics indicates a residue with positive phi values. Gray residues were not built in the model. Bracketed numbers refer to residues not depicted.

Article Snippet: Full-length and mutant Nup133 proteins (amino acids 1–1156, 1–514, and 497–1156) were in vitro transcribed and translated in the presence of [ 35 S]methionine by a coupled reticulocyte lysate transcription/translation system (TNT T7; Promega).

Techniques: Sequencing

Journal: The Journal of Cell Biology

Article Title: Structural and functional analysis of Nup133 domains reveals modular building blocks of the nuclear pore complex

doi: 10.1083/jcb.200408109

Figure Lengend Snippet: Conserved features of the Nup133 NTD propeller. (a) Surface representation showing conservation of residues on the propeller sides. Each view is an ∼120° rotation about the pseudo-sevenfold axis beginning from the NH 2 and COOH termini in blade 7 and is the same as in b. The tube marks the region of the disordered DA 34 loop. Conservation scores were calculated using the ConSurf Server ( http://consurf.tau.ac.il/ ) and colored maroon (most conserved) to cyan (most variable). (b) Electrostatic potential of the Nup133 β-propeller. Orientation is as in part a. Red indicates negative regions of the potential and blue indicates positive regions. (c) Sequence alignment of the DA 34 loop across representative metazoans. Position of the conserved protein kinase A consensus site is boxed and potential phosphorylated residues are marked by yellow stars. Green triangles mark conserved hydrophobic positions.

Article Snippet: Full-length and mutant Nup133 proteins (amino acids 1–1156, 1–514, and 497–1156) were in vitro transcribed and translated in the presence of [ 35 S]methionine by a coupled reticulocyte lysate transcription/translation system (TNT T7; Promega).

Techniques: Sequencing