rabbit α shc3  (Proteintech)

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

Journal: Journal of Hepatocellular Carcinoma

Article Title: The Long Non-Coding RNA AC006329.1 Facilitates Hepatocellular Carcinoma Progression and Metastasis by Regulating miR-127-5p/SHC3/ERK Axis

doi: 10.2147/JHC.S415309

Figure Lengend Snippet: SHC3 is a target gene of miR-127-5p. ( A ) Venn diagram analysis showed that SHC3 can be a target gene of miR-127-5p by four databases; ( B ) Volcano plot of differentially expressed mRNA in the TCGA-LIHC database. Red dots represent up-regulated mRNAs and blue dots represent down-regulated mRNAs in HCC tissues (threshold, p-value < 0.05 and |log2FoldChange| > 1); ( C ) Venn diagram was drawn to screen out mRNA with high expression in HCC tissues from TCGA database; ( D ) Pan-cancer analysis of the expression level of SHC3 in cancer tissues in TCGA database by Timer1.0; ( E ) Kaplan-Meier survival curve of the relationship between the expression of SHC3 and OS in HCC patients; ( F ) The expression of SHC3 in 50 pairs of HCC tissues and matched normal adjacent tissues via qRT-PCR; ( G ) The expression of SHC3 in 3 pairs of HCC and paracancer tissues was detected by WB assay; ( H ) Pearson correlation analysis was used to analyze the correlation between miR-127-5p and SHC3 in 50 pairs of HCC tissues; ( I ) Correlation analysis of the expression level of SHC3 and AC006329.1 in liver cancer tissues in TCGA database by GEPIA2; ( J ) The binding sites between miR-127-5p and SHC3 were predicted by starBase; ( K ) Luciferase activity was measured in HEK-293T cells cotransfected with SHC3-wt or SHC3-Mut luciferase plasmid and miR-127-5p mimic or mi-NC; ( L ) Spearman correlation analysis of SHC3 expression level and EMT marker expression (i), extracellular matrix related gene expression ( ii ) and extracellular matrix degradation level ( iii ) based on TCGA database; ( M ) Biocarta pathway enrichment analysis of AC006329.1 through starbase database; ( N ) Correlation analysis of expression levels of SHC3, ERK1 and ERK2 in TCGA database by GEPIA2. (** P < 0.01, *** P < 0.001,).

Article Snippet: The expression levels of Ki-67 and SHC3 were detected by immunohistochemical staining using specific antibodies against Ki-67 and SHC3 (Proteintech, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Luciferase, Activity Assay, Plasmid Preparation, Marker

Journal: Journal of Hepatocellular Carcinoma

Article Title: The Long Non-Coding RNA AC006329.1 Facilitates Hepatocellular Carcinoma Progression and Metastasis by Regulating miR-127-5p/SHC3/ERK Axis

doi: 10.2147/JHC.S415309

Figure Lengend Snippet: AC006329.1 promoted HCC by regulating the miR-127-5p/SHC3/ERK axis. ( A and B ) Colony formation assay showed that both mi-miR-127-5p and si-SHC3 reversed the enhancement of ov-AC006329.1 in HCC cell proliferation; ( C ) CCK-8 assay showed that both mi-miR-127-5p and si-SHC3 reversed the enhancement of ov-AC006329.1 in HCC cell proliferation; ( D and E ) EDU assay showed that both mi-miR-127-5p and si-SHC3 reversed the enhancement of ov-AC006329.1 in HCC cell proliferation; Scale bar represents 25 µm. ( F and G ) Wound healing assay showed that both mi-miR-127-5p and si-SHC3 reversed the enhancement of ov-AC006329.1 in HCC cell proliferation; Scale bar represents 100 µm. ( H – J ) 3D Spheroid Invasion assay and transwell assay showed that both mi-miR-127-5p and si-SHC3 reversed the enhancement of ov-AC006329.1 in HCC cell invasion and migration; Scale bar represents 50 µm. ( K ) The protein expression of E-cadherin, N-cadherin and vimentin was measured by EMT assay; Scale bar represents 25 µm. ( L ) The protein expression of E-cadherin, N-cadherin, vimentin, SHC3 and p-ERK was measured by and Western blotting assay in Hep-3B cells. (* P <0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: The expression levels of Ki-67 and SHC3 were detected by immunohistochemical staining using specific antibodies against Ki-67 and SHC3 (Proteintech, Wuhan, China).

Techniques: Colony Assay, CCK-8 Assay, EdU Assay, Wound Healing Assay, Invasion Assay, Transwell Assay, Migration, Expressing, Western Blot

Journal: Journal of Hepatocellular Carcinoma

Article Title: The Long Non-Coding RNA AC006329.1 Facilitates Hepatocellular Carcinoma Progression and Metastasis by Regulating miR-127-5p/SHC3/ERK Axis

doi: 10.2147/JHC.S415309

Figure Lengend Snippet: AC006329.1 promoted the progression and metastasis of HCC cells in vivo. ( A ) Photos of subcutaneous tumors in different groups of nude mice; Scale bar represents 100 µm. ( B ) Histogram of final weight of subcutaneous tumors in four groups of nude mice; ( C ) Volume growth curves of subcutaneous tumors in four groups of nude mice; ( D )The expression levels of Ki67 and SHC3 in subcutaneous tumor tissues of four groups detected by hematoxylin and eosin staining and immunohistochemistry; ( E ) H&E staining images of lung metastasis of nude mice injected with Hep-3B cells in the above four groups; Scale bar represents 100 µm; ( F ) The number of lung metastatic nodules in the above four groups. (** P < 0.01, *** P < 0.001).

Article Snippet: The expression levels of Ki-67 and SHC3 were detected by immunohistochemical staining using specific antibodies against Ki-67 and SHC3 (Proteintech, Wuhan, China).

Techniques: In Vivo, Expressing, Staining, Immunohistochemistry, Injection

Journal: Journal of Hepatocellular Carcinoma

Article Title: The Long Non-Coding RNA AC006329.1 Facilitates Hepatocellular Carcinoma Progression and Metastasis by Regulating miR-127-5p/SHC3/ERK Axis

doi: 10.2147/JHC.S415309

Figure Lengend Snippet: A simple pattern of targeted regulation of miR-127-5p/SHC3/ERK axis by AC006329.1 to promote the occurrence and development of HCC.

Article Snippet: The expression levels of Ki-67 and SHC3 were detected by immunohistochemical staining using specific antibodies against Ki-67 and SHC3 (Proteintech, Wuhan, China).

Techniques: