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GenePharma Company ribonucleic acid sirna
Specific downregulation of tripartite motif (TRIM)29 messenger ribonucleic acid (mRNA) and protein expression by <t>TRIM29</t> small interfering (si)RNA. Cells were transfected with TRIM29 <t>siRNA</t> or scrambled siRNA (siCONTROL) for 48 hours. (a) The relative mRNA level of TRIM29 was quantified by real-time polymerase chain reaction analysis, and 18S was used as an internal standard. (b) The relative TRIM29 protein level was determined by Western blotting, and data was normalized using β-actin as an internal standard. Compared with TRIM29 siRNA1 and 2, siRNA3 mostly silenced TRIM29 expression. P
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1) Product Images from "Silencing of tripartite motif (TRIM) 29 inhibits proliferation and invasion and increases chemosensitivity to cisplatin in human lung squamous cancer NCI-H520 cells"

Article Title: Silencing of tripartite motif (TRIM) 29 inhibits proliferation and invasion and increases chemosensitivity to cisplatin in human lung squamous cancer NCI-H520 cells

Journal: Thoracic Cancer

doi: 10.1111/1759-7714.12130

Specific downregulation of tripartite motif (TRIM)29 messenger ribonucleic acid (mRNA) and protein expression by TRIM29 small interfering (si)RNA. Cells were transfected with TRIM29 siRNA or scrambled siRNA (siCONTROL) for 48 hours. (a) The relative mRNA level of TRIM29 was quantified by real-time polymerase chain reaction analysis, and 18S was used as an internal standard. (b) The relative TRIM29 protein level was determined by Western blotting, and data was normalized using β-actin as an internal standard. Compared with TRIM29 siRNA1 and 2, siRNA3 mostly silenced TRIM29 expression. P
Figure Legend Snippet: Specific downregulation of tripartite motif (TRIM)29 messenger ribonucleic acid (mRNA) and protein expression by TRIM29 small interfering (si)RNA. Cells were transfected with TRIM29 siRNA or scrambled siRNA (siCONTROL) for 48 hours. (a) The relative mRNA level of TRIM29 was quantified by real-time polymerase chain reaction analysis, and 18S was used as an internal standard. (b) The relative TRIM29 protein level was determined by Western blotting, and data was normalized using β-actin as an internal standard. Compared with TRIM29 siRNA1 and 2, siRNA3 mostly silenced TRIM29 expression. P

Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot

Downregulation of tripartite motif (TRIM)29 inhibits invasion of NCI-H520 cells. (a) NCI-H520, siCONTROL and TRIM29 small interfering ribonucleic acid (siRNA) cells were seeded in the upper chamber in medium supplemented with five percent fetal bovine serum (FBS. After 24 hours, cells were fixed, stained, and counted (bar shows 100 μm). (b) The number of invasive cells in the TRIM29 siRNA treated group were significantly lower than in the NCI-H520 and siCONTROL groups ( P
Figure Legend Snippet: Downregulation of tripartite motif (TRIM)29 inhibits invasion of NCI-H520 cells. (a) NCI-H520, siCONTROL and TRIM29 small interfering ribonucleic acid (siRNA) cells were seeded in the upper chamber in medium supplemented with five percent fetal bovine serum (FBS. After 24 hours, cells were fixed, stained, and counted (bar shows 100 μm). (b) The number of invasive cells in the TRIM29 siRNA treated group were significantly lower than in the NCI-H520 and siCONTROL groups ( P

Techniques Used: Staining

RNA interference (RNAi)-mediated downregulation of tripartite motif (TRIM)29 reduces NCI-H520 cell proliferation. Cells treated with TRIM29 small interfering (si)RNA, scrambled control siRNA or with medium alone were plated on a 96-well plate. Cells were evaluated for proliferation at 24-hour intervals by MTT assay. Data were from three independent experiments. P
Figure Legend Snippet: RNA interference (RNAi)-mediated downregulation of tripartite motif (TRIM)29 reduces NCI-H520 cell proliferation. Cells treated with TRIM29 small interfering (si)RNA, scrambled control siRNA or with medium alone were plated on a 96-well plate. Cells were evaluated for proliferation at 24-hour intervals by MTT assay. Data were from three independent experiments. P

Techniques Used: MTT Assay

Small interfering ribonucleic acid (siRNA) for tripartite motif (TRIM29) induces apoptosis and enhances chemosensitivity. (a and c) TRIM29 inhibition by siRNA induced apoptosis in NCI-H520 cells. (b and d) After treating with 5 μg/mL cisplatin, cell apoptosis (including early and late apoptosis) was further enhanced. Combining TRIM29 inhibition with cisplatin increased the incidence of apoptosis. After 48 hours transfecting with siRNA, cells were treated with 5 μg/mL cisplatin for 24 hours, and then cells were double stained with Annexin V-FITC and propidium iodide (PI) followed by FACS analysis. FACS analysis scatter-grams of Annexin V/PI staining display four different cell populations marked as: double negative (unstained) cells showing live cell population (lower left), Annexin V positive and PI negative stained cells showing early apoptosis (lower right), Annexin V/PI double-stained cells showing late apoptosis (upper right), and, finally, PI positive and Annexin V negative stained cells showing dead cells (upper left). P
Figure Legend Snippet: Small interfering ribonucleic acid (siRNA) for tripartite motif (TRIM29) induces apoptosis and enhances chemosensitivity. (a and c) TRIM29 inhibition by siRNA induced apoptosis in NCI-H520 cells. (b and d) After treating with 5 μg/mL cisplatin, cell apoptosis (including early and late apoptosis) was further enhanced. Combining TRIM29 inhibition with cisplatin increased the incidence of apoptosis. After 48 hours transfecting with siRNA, cells were treated with 5 μg/mL cisplatin for 24 hours, and then cells were double stained with Annexin V-FITC and propidium iodide (PI) followed by FACS analysis. FACS analysis scatter-grams of Annexin V/PI staining display four different cell populations marked as: double negative (unstained) cells showing live cell population (lower left), Annexin V positive and PI negative stained cells showing early apoptosis (lower right), Annexin V/PI double-stained cells showing late apoptosis (upper right), and, finally, PI positive and Annexin V negative stained cells showing dead cells (upper left). P

Techniques Used: Inhibition, Staining, FACS

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Cell Culture:

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Polymerase Chain Reaction:

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Injection:

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In Vivo:

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Isolation:

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Mouse Assay:

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Small Interfering RNA:

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts
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Article Title: miR-382 Contributes to Renal Tubulointerstitial Fibrosis by Downregulating HSPD1
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Article Title: Npro of Classical Swine Fever Virus Suppresses Type III Interferon Production by Inhibiting IRF1 Expression and Its Nuclear Translocation
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Construct:

Article Title: Expression Analysis of Combinatorial Genes Using a Bi-Cistronic T2A Expression System in Porcine Fibroblasts
Article Snippet: To examine the protein expression pattern of these constructs, 2 or 3 µg of each plasmid were transiently introduced into 1×106 porcine fibroblasts. .. For RNA interference, we used 1 µg of small interfering RNA (siRNA) targeting GFP or luciferase GL2 (siGFP or siLuc, respectively; GenePharma, Shanghai, China).

Titration:

Article Title: Npro of Classical Swine Fever Virus Suppresses Type III Interferon Production by Inhibiting IRF1 Expression and Its Nuclear Translocation
Article Snippet: The anti-CSFV activity of IFN-λ3 (100 ng/mL) in IPEC-J2 cells were further examined 12 h before, at the time of, or after inoculation with wtCSFV or ∆Npro, and the whole cultures were collected at 24 hpi for virus titration. .. For gene silencing, IPEC-J2 cells were transfected with the small interfering RNA (siRNA) against IRF1 ( ) for 24 h with scrambled control siRNA (siNC) included as a control (Genepharma, Shanghai, China).

Plasmid Preparation:

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts
Article Snippet: .. Cell treatments To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation. .. Randomly scrambled siRNA or empty vector was transfected as a negative control respectively.

Article Title: Npro of Classical Swine Fever Virus Suppresses Type III Interferon Production by Inhibiting IRF1 Expression and Its Nuclear Translocation
Article Snippet: In order to examine modulation of transcription of IFN-λs or IRF1 by CSFV or Npro protein, IPEC-J2 cells were inoculated with wtCSFV or ∆Npro strain (MOI = 1) or transfected with the plasmid pCMV-Npro for 24 h, and then stimulated with 100 ng/mL poly(I:C) (Invivogen, USA) for 12 h. The cells were then lysed at various time points post-infection to analyze the transcription levels of IFN-λs or IRF1 by qRT-PCR using the primers listed in . .. For gene silencing, IPEC-J2 cells were transfected with the small interfering RNA (siRNA) against IRF1 ( ) for 24 h with scrambled control siRNA (siNC) included as a control (Genepharma, Shanghai, China).

Article Title: Expression Analysis of Combinatorial Genes Using a Bi-Cistronic T2A Expression System in Porcine Fibroblasts
Article Snippet: Paragraph title: Plasmid Construction and Transfection ... For RNA interference, we used 1 µg of small interfering RNA (siRNA) targeting GFP or luciferase GL2 (siGFP or siLuc, respectively; GenePharma, Shanghai, China).

Negative Control:

Article Title: Down-regulation of PDK4 is Critical for the Switch of Carbohydrate Catabolism during Syncytialization of Human Placental Trophoblasts
Article Snippet: Cell treatments To study the roles of PDK4 in the regulation of carbohydrate metabolism during syncytialization, the cells were transfected with small interfering RNA (siRNA) targeting PDK4 (sense: GAUGCUCUGUGAUCAGUAUTT, antisense: AUACUGAUCACAGAGCAUCTT) (GenePharma Co., Ltd., Shanghai, China) or a eukaryotic vector GV230 expressing PDK4 (GeneChem Co., Ltd. Shanghai, China) immediately after isolation. .. Randomly scrambled siRNA or empty vector was transfected as a negative control respectively.

Article Title: miR-382 Contributes to Renal Tubulointerstitial Fibrosis by Downregulating HSPD1
Article Snippet: .. In vivo delivery of small interfering RNA (siRNA) designed against HSPD1 or negative control siRNA (10 μ g, GenePharma) was performed by retrograde injection into the ureter proximal to the ligature, immediately following occlusion of the ureter. .. Mice were sacrificed by cervical dislocation 7 days after left ureteral obstruction.

Article Title: Silencing of tripartite motif (TRIM) 29 inhibits proliferation and invasion and increases chemosensitivity to cisplatin in human lung squamous cancer NCI-H520 cells
Article Snippet: The human TRIM29 specific small interfering ribonucleic acid (siRNA) was purchased from GenePharma Co Ltd (Shanghai, China). .. The sequences of the siRNA used were as follows: siRNA1: sense, 5′-GAGCUGCGCAAGUCCAUUUTT-3′, siRNA2: sense,5′-ACGGAGCUGUCAUUGCAAATT-3′, siRNA3:sense, 5′-GUGCAUUGAUGAGCAAUUATT-3′, Negative control siRNA: sense, 5′-UUCUCCGAACGUGUCACGUTT-3′.

Article Title: Circular RNA hsa-circ-0012129 Promotes Cell Proliferation and Invasion in 30 Cases of Human Glioma and Human Glioma Cell Lines U373, A172, and SHG44, by Targeting MicroRNA-661 (miR-661)
Article Snippet: .. Cell transfection with small interfering RNA (siRNA) For transfection with small interfering RNA (siRNA), negative control (NC) siRNAs, and siRNAs for hsa-circ-0012129 were obtained from GenePharma Co., Ltd (Shanghai, China). .. One day before transfection, U373 and SHG44 cells (2×104 cells/well) in the logarithmic phase were seeded into six-well plates in 2 mL of medium and incubated overnight.

Produced:

Article Title: miR-382 Contributes to Renal Tubulointerstitial Fibrosis by Downregulating HSPD1
Article Snippet: The obstruction was produced by ligation of the left ureter following midline laparotomy. .. In vivo delivery of small interfering RNA (siRNA) designed against HSPD1 or negative control siRNA (10 μ g, GenePharma) was performed by retrograde injection into the ureter proximal to the ligature, immediately following occlusion of the ureter.

Staining:

Article Title: miR-382 Contributes to Renal Tubulointerstitial Fibrosis by Downregulating HSPD1
Article Snippet: In vivo delivery of small interfering RNA (siRNA) designed against HSPD1 or negative control siRNA (10 μ g, GenePharma) was performed by retrograde injection into the ureter proximal to the ligature, immediately following occlusion of the ureter. .. The degree of renal fibrosis was graded according to Masson trichrome staining and Sirius red staining.

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  • 94
    GenePharma Company drp1 sirna
    Tan IIA-mediated mitochondrial fission triggers mitochondrial damage. a-b Mitochondrial ROS production was measured via flow cytometry. Mdivi-1 and <t>Drp1</t> <t>siRNA</t> were used to prevent fission, which was used as the loss-of-function assay for mitochondrial fission. The mitochondrial fission agonist (FCCP) was added to the control group to activate mitochondrial fission, and this was used to mimic the effects of Tan IIA. c-d Antioxidant factors, including SOD and GPX, were quantified via ELISA in SW837 cells treated with Tan IIA. e-f Immunofluorescence assay for Smac. Blue fluorescence is from the DAPI probe, which stains the nucleus. g Caspase-9 activity was determined via ELISA. Mdivi-1 and Drp1 siRNA were used to prevent fission, which was used as the loss-of-function assay for mitochondrial fission. The mitochondrial fission agonist (FCCP) was added to the control group to activate mitochondrial fission, and this was used to mimic the effects of Tan IIA. h-i TUNEL staining to assess cell death. The number of TUNEL-positive cell was counted. * p
    Drp1 Sirna, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/drp1 sirna/product/GenePharma Company
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    93
    GenePharma Company sirna controls
    <t>STAT3</t> mediates EMT reversal induced by YM155. a Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705), STAT3, E-cadherin, and N-cadherin in U251 and U87 cells before and after knockdown with STAT3 <t>siRNA.</t> b Morphology of U251 and U87 cells before and after STAT3 siRNA knockdown. c Crystal violet staining of Transwell Matrigel assay for U251 and U87 cells control or transfected with STAT3 siRNA after radiation treatment. d Quantitation of migrating cell number in experiments from ( c ). e Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705) and STAT3 in lysates prepared from control cells and cells stably expressing STAT3. f Crystal violet staining of Transwell Matrigel assay from control or stable ectopic expression of STAT3 U251 cells after control, radiation (4 Gy), YM155 (5 nM) and combination treatment (5 nM YM155 + 4 Gy radiation). g Quantitation of migrating cell number in experiments from ( f ). h Crystal violet staining of Transwell Matrigel assay from YM155 treated U251 and U87 cells or stable ectopic expression of STAT3 U251 and U87 cells following YM155 treatment. i Quantitation of migrating cell number in experiments from ( h ). ** P
    Sirna Controls, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 93/100, based on 186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    GenePharma Company silencing stathmin shrna
    Suppression of <t>Stathmin</t> in hDPSCs inhibited proliferation and the expression of mineralization‐related genes in hDPSCs. A‐C, Cell cycle distribution for the <t>shRNA‐Ctrl</t> group (A) and the shRNA‐Stathmin group (B) and statistical analysis (C). D, CCK‐8 value of shRNA‐Stathmin hDPSCs and shRNA‐Ctrl hDPSCs at days 1, 3, 5 and 7. Mineralization‐related genes (E, ALP; F, BSP; G, OCN; H, DSPP) were expressed at much lower levels in the Stathmin knockdown group than in the shRNA‐Ctrl group cultured in mineralization medium for 3 weeks. I and J, Alizarin red S staining showed that Stathmin suppression significantly inhibited mineral formation by hDPSCs (scale bar = 100 μm). Each experiment was repeated in triplicate. (* P
    Silencing Stathmin Shrna, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Tan IIA-mediated mitochondrial fission triggers mitochondrial damage. a-b Mitochondrial ROS production was measured via flow cytometry. Mdivi-1 and Drp1 siRNA were used to prevent fission, which was used as the loss-of-function assay for mitochondrial fission. The mitochondrial fission agonist (FCCP) was added to the control group to activate mitochondrial fission, and this was used to mimic the effects of Tan IIA. c-d Antioxidant factors, including SOD and GPX, were quantified via ELISA in SW837 cells treated with Tan IIA. e-f Immunofluorescence assay for Smac. Blue fluorescence is from the DAPI probe, which stains the nucleus. g Caspase-9 activity was determined via ELISA. Mdivi-1 and Drp1 siRNA were used to prevent fission, which was used as the loss-of-function assay for mitochondrial fission. The mitochondrial fission agonist (FCCP) was added to the control group to activate mitochondrial fission, and this was used to mimic the effects of Tan IIA. h-i TUNEL staining to assess cell death. The number of TUNEL-positive cell was counted. * p

    Journal: BMC Cell Biology

    Article Title: Tanshinone IIA reduces SW837 colorectal cancer cell viability via the promotion of mitochondrial fission by activating JNK-Mff signaling pathways

    doi: 10.1186/s12860-018-0174-z

    Figure Lengend Snippet: Tan IIA-mediated mitochondrial fission triggers mitochondrial damage. a-b Mitochondrial ROS production was measured via flow cytometry. Mdivi-1 and Drp1 siRNA were used to prevent fission, which was used as the loss-of-function assay for mitochondrial fission. The mitochondrial fission agonist (FCCP) was added to the control group to activate mitochondrial fission, and this was used to mimic the effects of Tan IIA. c-d Antioxidant factors, including SOD and GPX, were quantified via ELISA in SW837 cells treated with Tan IIA. e-f Immunofluorescence assay for Smac. Blue fluorescence is from the DAPI probe, which stains the nucleus. g Caspase-9 activity was determined via ELISA. Mdivi-1 and Drp1 siRNA were used to prevent fission, which was used as the loss-of-function assay for mitochondrial fission. The mitochondrial fission agonist (FCCP) was added to the control group to activate mitochondrial fission, and this was used to mimic the effects of Tan IIA. h-i TUNEL staining to assess cell death. The number of TUNEL-positive cell was counted. * p

    Article Snippet: The sense and antisense strands of the Drp1 siRNA were 5′-GGCAGAGGAAGAAUAUAAATT-3′ and 5′-UUUAUAUUCUUCCUCUGCCTT-3′; negative control were 5′-UUCUCCGAACGUGUCACGUTT-3′ and 5′-ACGUGACACGUUCGGAGAATT-3′. (designed and synthesized by GenePharma Co. Ltd., Shanghai).

    Techniques: Flow Cytometry, Cytometry, Functional Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Fluorescence, Activity Assay, TUNEL Assay, Staining

    STAT3 mediates EMT reversal induced by YM155. a Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705), STAT3, E-cadherin, and N-cadherin in U251 and U87 cells before and after knockdown with STAT3 siRNA. b Morphology of U251 and U87 cells before and after STAT3 siRNA knockdown. c Crystal violet staining of Transwell Matrigel assay for U251 and U87 cells control or transfected with STAT3 siRNA after radiation treatment. d Quantitation of migrating cell number in experiments from ( c ). e Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705) and STAT3 in lysates prepared from control cells and cells stably expressing STAT3. f Crystal violet staining of Transwell Matrigel assay from control or stable ectopic expression of STAT3 U251 cells after control, radiation (4 Gy), YM155 (5 nM) and combination treatment (5 nM YM155 + 4 Gy radiation). g Quantitation of migrating cell number in experiments from ( f ). h Crystal violet staining of Transwell Matrigel assay from YM155 treated U251 and U87 cells or stable ectopic expression of STAT3 U251 and U87 cells following YM155 treatment. i Quantitation of migrating cell number in experiments from ( h ). ** P

    Journal: Journal of Translational Medicine

    Article Title: YM155 decreases radiation-induced invasion and reverses epithelial–mesenchymal transition by targeting STAT3 in glioblastoma

    doi: 10.1186/s12967-018-1451-5

    Figure Lengend Snippet: STAT3 mediates EMT reversal induced by YM155. a Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705), STAT3, E-cadherin, and N-cadherin in U251 and U87 cells before and after knockdown with STAT3 siRNA. b Morphology of U251 and U87 cells before and after STAT3 siRNA knockdown. c Crystal violet staining of Transwell Matrigel assay for U251 and U87 cells control or transfected with STAT3 siRNA after radiation treatment. d Quantitation of migrating cell number in experiments from ( c ). e Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705) and STAT3 in lysates prepared from control cells and cells stably expressing STAT3. f Crystal violet staining of Transwell Matrigel assay from control or stable ectopic expression of STAT3 U251 cells after control, radiation (4 Gy), YM155 (5 nM) and combination treatment (5 nM YM155 + 4 Gy radiation). g Quantitation of migrating cell number in experiments from ( f ). h Crystal violet staining of Transwell Matrigel assay from YM155 treated U251 and U87 cells or stable ectopic expression of STAT3 U251 and U87 cells following YM155 treatment. i Quantitation of migrating cell number in experiments from ( h ). ** P

    Article Snippet: Sequences for the siRNAs used were the following: survivin, 5′-GCATTCGTCCGGTTGCGCT-3′; STAT3, 5′-GUUCAUCUGUGUGACACCATT-3′; nontargeting siRNA controls, 5′-UUCUCCGAACGUGUCACGUTT-3′ (Genepharma, Shanghai, China).

    Techniques: Western Blot, Staining, Matrigel Assay, Transfection, Quantitation Assay, Stable Transfection, Expressing

    STAT3 mediates EMT reversal induced by YM155 independently of survivin. a Morphology of U251 and U87 cells before and after knockdown with survivin siRNA, or knockdown with YM155 treatment. b Phospho-kinase array of 43 phosphorylated kinases in U251 cells treated with 5 nM YM155 relative to DMSO control. c Graphic representation of the quantitation of 10 molecules with the most significant difference in phosphorylation status between YM155 treatment and control. d Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705), STAT3, Cyclin D1 and c-Myc of U251 and U87 cells incubated with increasing concentrations of YM155. e Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705), STAT3, N-cadherin and E-cadherin in U251 and U87 cells treated with DMSO, radiation, and combination treatment (YM155 5 nM + radiation 4 Gy)

    Journal: Journal of Translational Medicine

    Article Title: YM155 decreases radiation-induced invasion and reverses epithelial–mesenchymal transition by targeting STAT3 in glioblastoma

    doi: 10.1186/s12967-018-1451-5

    Figure Lengend Snippet: STAT3 mediates EMT reversal induced by YM155 independently of survivin. a Morphology of U251 and U87 cells before and after knockdown with survivin siRNA, or knockdown with YM155 treatment. b Phospho-kinase array of 43 phosphorylated kinases in U251 cells treated with 5 nM YM155 relative to DMSO control. c Graphic representation of the quantitation of 10 molecules with the most significant difference in phosphorylation status between YM155 treatment and control. d Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705), STAT3, Cyclin D1 and c-Myc of U251 and U87 cells incubated with increasing concentrations of YM155. e Western blot analysis of pSTAT3 (Ser727), pSTAT3 (Tyr705), STAT3, N-cadherin and E-cadherin in U251 and U87 cells treated with DMSO, radiation, and combination treatment (YM155 5 nM + radiation 4 Gy)

    Article Snippet: Sequences for the siRNAs used were the following: survivin, 5′-GCATTCGTCCGGTTGCGCT-3′; STAT3, 5′-GUUCAUCUGUGUGACACCATT-3′; nontargeting siRNA controls, 5′-UUCUCCGAACGUGUCACGUTT-3′ (Genepharma, Shanghai, China).

    Techniques: Quantitation Assay, Western Blot, Incubation

    Knockdown of MALAT1 inhibits ox-LDL-induced β-catenin translocation. HUVECs were transfected with MALAT1-siRNA or scramble control (scr) for 24 h. a The protein expression of β-catenin in nucleus (n = 3, * P

    Journal: Lipids in Health and Disease

    Article Title: LncRNA MALAT1 modulates ox-LDL induced EndMT through the Wnt/β-catenin signaling pathway

    doi: 10.1186/s12944-019-1006-7

    Figure Lengend Snippet: Knockdown of MALAT1 inhibits ox-LDL-induced β-catenin translocation. HUVECs were transfected with MALAT1-siRNA or scramble control (scr) for 24 h. a The protein expression of β-catenin in nucleus (n = 3, * P

    Article Snippet: For MALAT1 knockdown, three MALAT1-targeting siRNAs (MALAT1-siRNA1, 5′-GGUGGUGG UAUUUAGAUAATTUUAUCUAAA UACCACCACCTT-3′; MALAT1-siRNA2, 5′-GCGUCAUUUAAAGCCUAGUTTA CUAGGCUUUAAAUGACGCTT3’; MALAT1-siRNA3, 5′-GGGCUGACAUUAAC UACAATTUUGUAGUUAAUGUCA GCCCTT-3′) and a scrambled siRNA (sense 5′-UUCUCCGAACGUGUCACGUTT-3′, antisense 5′-ACGUGACACGUUCGGAGAA TT-3′) (Scramble) were designed and synthesized at GenePharma (Shanghai, China).

    Techniques: Translocation Assay, Transfection, Expressing

    Suppression of Stathmin in hDPSCs inhibited proliferation and the expression of mineralization‐related genes in hDPSCs. A‐C, Cell cycle distribution for the shRNA‐Ctrl group (A) and the shRNA‐Stathmin group (B) and statistical analysis (C). D, CCK‐8 value of shRNA‐Stathmin hDPSCs and shRNA‐Ctrl hDPSCs at days 1, 3, 5 and 7. Mineralization‐related genes (E, ALP; F, BSP; G, OCN; H, DSPP) were expressed at much lower levels in the Stathmin knockdown group than in the shRNA‐Ctrl group cultured in mineralization medium for 3 weeks. I and J, Alizarin red S staining showed that Stathmin suppression significantly inhibited mineral formation by hDPSCs (scale bar = 100 μm). Each experiment was repeated in triplicate. (* P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Stathmin inhibits proliferation and differentiation of dental pulp stem cells via sonic hedgehog/Gli, et al. Stathmin inhibits proliferation and differentiation of dental pulp stem cells via sonic hedgehog/Gli

    doi: 10.1111/jcmm.13621

    Figure Lengend Snippet: Suppression of Stathmin in hDPSCs inhibited proliferation and the expression of mineralization‐related genes in hDPSCs. A‐C, Cell cycle distribution for the shRNA‐Ctrl group (A) and the shRNA‐Stathmin group (B) and statistical analysis (C). D, CCK‐8 value of shRNA‐Stathmin hDPSCs and shRNA‐Ctrl hDPSCs at days 1, 3, 5 and 7. Mineralization‐related genes (E, ALP; F, BSP; G, OCN; H, DSPP) were expressed at much lower levels in the Stathmin knockdown group than in the shRNA‐Ctrl group cultured in mineralization medium for 3 weeks. I and J, Alizarin red S staining showed that Stathmin suppression significantly inhibited mineral formation by hDPSCs (scale bar = 100 μm). Each experiment was repeated in triplicate. (* P

    Article Snippet: 2.3 Lentiviral vector cell transduction A lentivirus containing silencing Stathmin shRNA and the green fluorescent protein (GFP) gene and a negative control consisting of a scrambled sequence and GFP were constructed by the GenePharma Company (GenePharma, Shanghai, China). hDPSCs were infected with shRNA‐Stathmin and shRNA‐Ctrl lentiviruses at a multiplicity of infection of 50.

    Techniques: Expressing, shRNA, CCK-8 Assay, ALP Assay, Cell Culture, Staining

    Expression of Stathmin in hDPSCs and lentivirus infections of hDPSCs. A, B, Expression of Stathmin in the cytomembrane and cytoplasm of hDPSCs (scale bar = 20 μm). C, D, At 72 h after gene transduction, both shRNA‐Stathmin‐ and shRNA‐Ctrl‐transfected hDPSCs grew well, and green fluorescent protein (GFP) fluorescence in the hDPSCs confirmed the transfection efficiency (scale bar = 100 μm). E, F, Western blot analysis showing that the Stathmin protein band in the shRNA‐Stathmin group was significantly weaker than that in the shRNA‐Ctrl group. G, Real‐time PCR showed that the expression levels of Stathmin messenger RNA were lower in the shRNA‐Ctrl group than in the shRNA‐Stathmin group ( ***P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Stathmin inhibits proliferation and differentiation of dental pulp stem cells via sonic hedgehog/Gli, et al. Stathmin inhibits proliferation and differentiation of dental pulp stem cells via sonic hedgehog/Gli

    doi: 10.1111/jcmm.13621

    Figure Lengend Snippet: Expression of Stathmin in hDPSCs and lentivirus infections of hDPSCs. A, B, Expression of Stathmin in the cytomembrane and cytoplasm of hDPSCs (scale bar = 20 μm). C, D, At 72 h after gene transduction, both shRNA‐Stathmin‐ and shRNA‐Ctrl‐transfected hDPSCs grew well, and green fluorescent protein (GFP) fluorescence in the hDPSCs confirmed the transfection efficiency (scale bar = 100 μm). E, F, Western blot analysis showing that the Stathmin protein band in the shRNA‐Stathmin group was significantly weaker than that in the shRNA‐Ctrl group. G, Real‐time PCR showed that the expression levels of Stathmin messenger RNA were lower in the shRNA‐Ctrl group than in the shRNA‐Stathmin group ( ***P

    Article Snippet: 2.3 Lentiviral vector cell transduction A lentivirus containing silencing Stathmin shRNA and the green fluorescent protein (GFP) gene and a negative control consisting of a scrambled sequence and GFP were constructed by the GenePharma Company (GenePharma, Shanghai, China). hDPSCs were infected with shRNA‐Stathmin and shRNA‐Ctrl lentiviruses at a multiplicity of infection of 50.

    Techniques: Expressing, Transduction, shRNA, Transfection, Fluorescence, Western Blot, Real-time Polymerase Chain Reaction

    Activation of Shh signalling promotes the proliferation and osteogenic/odontoblastic differentiation of hDPSCs. A‐D, Treatment of shRNA‐Stathmin hDPSCs with purmorphamine that specifically binds SMO increased (A, ALP; B, BSP; C, OCN; D, DSPP) mRNA expression, as determined by real‐time PCR. E‐G, Cell cycle distribution for the shRNA‐Stathmin group (E) and the shRNA‐Stathmin + PM group (F) and statistical analysis (G). H, CCK‐8 values of shRNA‐Stathmin + PM hDPSCs and shRNA‐Stathmin hDPSCs at days 1, 3, 5 and 7. Each experiment was repeated in triplicate (* P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Stathmin inhibits proliferation and differentiation of dental pulp stem cells via sonic hedgehog/Gli, et al. Stathmin inhibits proliferation and differentiation of dental pulp stem cells via sonic hedgehog/Gli

    doi: 10.1111/jcmm.13621

    Figure Lengend Snippet: Activation of Shh signalling promotes the proliferation and osteogenic/odontoblastic differentiation of hDPSCs. A‐D, Treatment of shRNA‐Stathmin hDPSCs with purmorphamine that specifically binds SMO increased (A, ALP; B, BSP; C, OCN; D, DSPP) mRNA expression, as determined by real‐time PCR. E‐G, Cell cycle distribution for the shRNA‐Stathmin group (E) and the shRNA‐Stathmin + PM group (F) and statistical analysis (G). H, CCK‐8 values of shRNA‐Stathmin + PM hDPSCs and shRNA‐Stathmin hDPSCs at days 1, 3, 5 and 7. Each experiment was repeated in triplicate (* P

    Article Snippet: 2.3 Lentiviral vector cell transduction A lentivirus containing silencing Stathmin shRNA and the green fluorescent protein (GFP) gene and a negative control consisting of a scrambled sequence and GFP were constructed by the GenePharma Company (GenePharma, Shanghai, China). hDPSCs were infected with shRNA‐Stathmin and shRNA‐Ctrl lentiviruses at a multiplicity of infection of 50.

    Techniques: Activation Assay, shRNA, ALP Assay, Expressing, Real-time Polymerase Chain Reaction, CCK-8 Assay