Journal: PLOS ONE

Article Title: Short communication: Upregulation of hypoxia/reoxygenation-induced Shc3 by downregulated miR-455-5p, suppresses trophoblast invasion and is associated with placental inflammation and angiogenesis in preeclampsia

doi: 10.1371/journal.pone.0314544

Figure Lengend Snippet: (A) Heatmap of RNA-Seqs. (B) Selection of potential miR-455-5p downstream genes. (C) Schematic diagram of luciferase reporter-Shc3 3’UTR constructs. (D) The outcomes of miR-455-5p overexpression on the luciferase activity in HTR8/SVneo cells transfected with the WT or MUT luciferase reporter constructs. (E) The effect of miR-455-5p inhibition and overexpression on Shc3 mRNA expression in HTR8/SVneo cells. (F) The level of Shc3 protein expression after miR-455-5p inhibition and overexpression in HTR8/SVneo. Data presented shows the average values ± standard deviation (**p < 0.01, *p < 0.05). The findings presented were obtained from a minimum of three independent experiments.

Article Snippet: SHC3, Beta Actin antibodies and HRP-conjugated Affinipure Goat Anti Rabbit IgG(H+L) were obtained from Proteintech (Wuhan, China).

Techniques: Selection, Luciferase, Construct, Over Expression, Activity Assay, Transfection, Inhibition, Expressing, Standard Deviation

Journal: PLOS ONE

Article Title: Short communication: Upregulation of hypoxia/reoxygenation-induced Shc3 by downregulated miR-455-5p, suppresses trophoblast invasion and is associated with placental inflammation and angiogenesis in preeclampsia

doi: 10.1371/journal.pone.0314544

Figure Lengend Snippet: (A) The migration and invasion of HTR8/SVneo cells with miR-455-5p overexpression plasmids co-cultured with the plasmids of Shc3, or Shc3-NC in H/R. (B) Flow cytometry data of the apoptosis of HTR8/SVneo cells with miR-455-5p overexpression plasmids co-cultured with the plasmids expressing Shc3 or Shc3-NC in H/R. (C) Western blot results of Shc3 expression in HTR8/SVneo cells transfected with the miR-455-5p mimic, Shc3, or Shc3-NC. (D) The results of Shc3 expression examined by immunohistochemistry. Data presented indicate the average values ± standard deviation (**p < 0.01, *p < 0.05). The findings presented were obtained from a minimum of three independent experiments.

Article Snippet: SHC3, Beta Actin antibodies and HRP-conjugated Affinipure Goat Anti Rabbit IgG(H+L) were obtained from Proteintech (Wuhan, China).

Techniques: Migration, Over Expression, Cell Culture, Flow Cytometry, Expressing, Western Blot, Transfection, Immunohistochemistry, Standard Deviation

Journal: PLOS ONE

Article Title: Short communication: Upregulation of hypoxia/reoxygenation-induced Shc3 by downregulated miR-455-5p, suppresses trophoblast invasion and is associated with placental inflammation and angiogenesis in preeclampsia

doi: 10.1371/journal.pone.0314544

Figure Lengend Snippet: (A~B). H/R promotes the production of Shc3, which may be transported to the extracellular space through medium/large extracellular vesicles. (A) Western blot results of Shc3 expression in hypoxia, H/R, and normoxia, respectively. (B) Volcano plot of Shc3 mRNA expression in placental tissue, extracellular medium/large vesicles and small vesicles. (C~E). Shc3 is involved in placental inflammation and angiogenesis inhibition. (C) Hematoxylin-eosin staining results on normotensive placental tissues and high Shc3 expression PE placental tissues. (D) The level of Shc3 in EA.hy926 after transfection with the plasmids of Shc3, and Shc3-NC, in H/R compared with controls. (E) Angiogenesis of EA.hy926 after transfection with the plasmids of Shc3, and Shc3-NC in H/R, as determined by the number of master junction and master segment length.

Article Snippet: SHC3, Beta Actin antibodies and HRP-conjugated Affinipure Goat Anti Rabbit IgG(H+L) were obtained from Proteintech (Wuhan, China).

Techniques: Western Blot, Expressing, Inhibition, Staining, Transfection