pex5 sirna Search Results


pex5 sirna #1  (Bioneer Corporation)
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Bioneer Corporation pex5 sirna #1
a HepG2 cells were treated with CTL <t>siRNA</t> (100 nM) and PEX5 siRNA #1 or #2 (100 nM) for 24 h. Then, cells were incubated in the presence or absence of serum. After 24 h, cells were harvested and immunoblotted with antibodies against PEX5, LC3I/II, p62, and β-actin. b Immunoblot analysis of p-70S6K, p-S6R, and p-4E-BP-1, and total 70S6K, S6R, and 4E-BP-1. c Cells <t>were</t> <t>transfected</t> with GFP-LC3 plasmid. After 6 h of transfection, cells were treated with CTL and PEX5 siRNAs. Visualization and quantification of GFP-LC3 puncta were performed by fluorescence microscopy after 48 h of incubation. Representative images from three independent determinations are shown (* P < 0.05). Scale bar: 10 μm. d Immunoblot analysis of TSC2 and PEX5 protein levels. e Transcript levels of TSC2 were measured by Q-PCR (** P < 0.01)
Pex5 Sirna #1, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a HepG2 cells were treated with CTL siRNA (100 nM) and PEX5 siRNA #1 or #2 (100 nM) for 24 h. Then, cells were incubated in the presence or absence of serum. After 24 h, cells were harvested and immunoblotted with antibodies against PEX5, LC3I/II, p62, and β-actin. b Immunoblot analysis of p-70S6K, p-S6R, and p-4E-BP-1, and total 70S6K, S6R, and 4E-BP-1. c Cells were transfected with GFP-LC3 plasmid. After 6 h of transfection, cells were treated with CTL and PEX5 siRNAs. Visualization and quantification of GFP-LC3 puncta were performed by fluorescence microscopy after 48 h of incubation. Representative images from three independent determinations are shown (* P < 0.05). Scale bar: 10 μm. d Immunoblot analysis of TSC2 and PEX5 protein levels. e Transcript levels of TSC2 were measured by Q-PCR (** P < 0.01)

Journal: Experimental & Molecular Medicine

Article Title: PEX5 regulates autophagy via the mTORC1-TFEB axis during starvation

doi: 10.1038/s12276-017-0007-8

Figure Lengend Snippet: a HepG2 cells were treated with CTL siRNA (100 nM) and PEX5 siRNA #1 or #2 (100 nM) for 24 h. Then, cells were incubated in the presence or absence of serum. After 24 h, cells were harvested and immunoblotted with antibodies against PEX5, LC3I/II, p62, and β-actin. b Immunoblot analysis of p-70S6K, p-S6R, and p-4E-BP-1, and total 70S6K, S6R, and 4E-BP-1. c Cells were transfected with GFP-LC3 plasmid. After 6 h of transfection, cells were treated with CTL and PEX5 siRNAs. Visualization and quantification of GFP-LC3 puncta were performed by fluorescence microscopy after 48 h of incubation. Representative images from three independent determinations are shown (* P < 0.05). Scale bar: 10 μm. d Immunoblot analysis of TSC2 and PEX5 protein levels. e Transcript levels of TSC2 were measured by Q-PCR (** P < 0.01)

Article Snippet: They were then transfected with 100 nM control siRNA, PEX5 siRNA (#1 sense-CGUAUCCUGGGAUCUCUCU, antisense-AGAGAGAUCCCAGGAUACG, #2 sense-GAAUUCAUCUCUGAAGUUA, antisense-UAACUUCAGAGAGAUGAAUUC-3, Bioneer, Daejeon, Korea), or TFEB siRNA (sc38509, Santa Cruz Biotechnology) for 24 h in serum-free medium, using Lipofectamine RNAiMax transfection reagent (Invitrogen/Life Technologies), and then exposed to serum starvation for 24 h. The effect of gene silencing was determined by Q-PCR and western blot analysis.

Techniques: Incubation, Western Blot, Transfection, Plasmid Preparation, Fluorescence, Microscopy

a Cells were treated with CTL and PEX5 siRNAs (100 nM) for 24 h. Then, cells were incubated in the presence or absence of serum. After 24 h, cells were stained with an antibody against TFEB and analyzed by fluorescence microscopy. Scale bar: 10 μm. b After HepG2 cells were treated as indicated, cells were subjected to nuclear and cytosol fractionation and immunoblotted with antibodies against TFEB, PCNA, and β-actin. PCNA and β-actin were used as nuclear and cytosolic markers, respectively. c , d Cells were transfected with CTL or TFEB siRNA (100 nM) for 24 h, and then incubated in the presence or absence of serum for 24 h. Protein expression of TFEB, LC3 and SQSTM1/p62 (autophagy proteins), and PMP70, DBP, and catalase (peroxisomal proteins) were measured by immunoblotting. e Q-PCR analysis of peroxisomal genes was performed (** P < 0.01, * P < 0.05)

Journal: Experimental & Molecular Medicine

Article Title: PEX5 regulates autophagy via the mTORC1-TFEB axis during starvation

doi: 10.1038/s12276-017-0007-8

Figure Lengend Snippet: a Cells were treated with CTL and PEX5 siRNAs (100 nM) for 24 h. Then, cells were incubated in the presence or absence of serum. After 24 h, cells were stained with an antibody against TFEB and analyzed by fluorescence microscopy. Scale bar: 10 μm. b After HepG2 cells were treated as indicated, cells were subjected to nuclear and cytosol fractionation and immunoblotted with antibodies against TFEB, PCNA, and β-actin. PCNA and β-actin were used as nuclear and cytosolic markers, respectively. c , d Cells were transfected with CTL or TFEB siRNA (100 nM) for 24 h, and then incubated in the presence or absence of serum for 24 h. Protein expression of TFEB, LC3 and SQSTM1/p62 (autophagy proteins), and PMP70, DBP, and catalase (peroxisomal proteins) were measured by immunoblotting. e Q-PCR analysis of peroxisomal genes was performed (** P < 0.01, * P < 0.05)

Article Snippet: They were then transfected with 100 nM control siRNA, PEX5 siRNA (#1 sense-CGUAUCCUGGGAUCUCUCU, antisense-AGAGAGAUCCCAGGAUACG, #2 sense-GAAUUCAUCUCUGAAGUUA, antisense-UAACUUCAGAGAGAUGAAUUC-3, Bioneer, Daejeon, Korea), or TFEB siRNA (sc38509, Santa Cruz Biotechnology) for 24 h in serum-free medium, using Lipofectamine RNAiMax transfection reagent (Invitrogen/Life Technologies), and then exposed to serum starvation for 24 h. The effect of gene silencing was determined by Q-PCR and western blot analysis.

Techniques: Incubation, Staining, Fluorescence, Microscopy, Fractionation, Transfection, Expressing, Western Blot

Cells were transfected with CTL siRNA (100 nM) and PEX5 siRNA (100 nM) for 24 h, and then incubated in the presence or absence of serum for 24 h. After incubation, cells were stained with LysoTracker Red DND-99 (50 nM) for 3 h and analyzed by confocal microscopy ( a ) or fluorescence-activated cell sorting and quantified ( b ). Data are presented as mean ± s.d. from three independent experiments (* P < 0.05). c Transcript levels of TFEB target genes were measured by Q-PCR. Values are expressed as fold increase compared to the starvation control group. Data were presented as mean ± s.d. from three independent experiments (** P < 0.01, * P < 0.05). d , e Protein expression of LAMP-1/LAMP-2 (lysosomal proteins) and cathepsins B/D (lysosomal proteases) were measured by immunoblotting

Journal: Experimental & Molecular Medicine

Article Title: PEX5 regulates autophagy via the mTORC1-TFEB axis during starvation

doi: 10.1038/s12276-017-0007-8

Figure Lengend Snippet: Cells were transfected with CTL siRNA (100 nM) and PEX5 siRNA (100 nM) for 24 h, and then incubated in the presence or absence of serum for 24 h. After incubation, cells were stained with LysoTracker Red DND-99 (50 nM) for 3 h and analyzed by confocal microscopy ( a ) or fluorescence-activated cell sorting and quantified ( b ). Data are presented as mean ± s.d. from three independent experiments (* P < 0.05). c Transcript levels of TFEB target genes were measured by Q-PCR. Values are expressed as fold increase compared to the starvation control group. Data were presented as mean ± s.d. from three independent experiments (** P < 0.01, * P < 0.05). d , e Protein expression of LAMP-1/LAMP-2 (lysosomal proteins) and cathepsins B/D (lysosomal proteases) were measured by immunoblotting

Article Snippet: They were then transfected with 100 nM control siRNA, PEX5 siRNA (#1 sense-CGUAUCCUGGGAUCUCUCU, antisense-AGAGAGAUCCCAGGAUACG, #2 sense-GAAUUCAUCUCUGAAGUUA, antisense-UAACUUCAGAGAGAUGAAUUC-3, Bioneer, Daejeon, Korea), or TFEB siRNA (sc38509, Santa Cruz Biotechnology) for 24 h in serum-free medium, using Lipofectamine RNAiMax transfection reagent (Invitrogen/Life Technologies), and then exposed to serum starvation for 24 h. The effect of gene silencing was determined by Q-PCR and western blot analysis.

Techniques: Transfection, Incubation, Staining, Confocal Microscopy, Fluorescence, FACS, Control, Expressing, Western Blot

Cells transfected with CTL siRNA (100 nM) and PEX5 siRNA (100 nM) for 24 h were pretreated with DMSO or rapamycin (1 μM) for 6 h, and then incubated in the absence of serum for 24 h. ( a ) After HepG2 cells were treated as indicated, cells were stained with an antibody against TFEB and analyzed by fluorescence microscopy. Scale bar: 10 μm. Data are presented as mean ± SD from three independent experiments (** P < 0.01). ( b ) After HepG2 cells were treated as indicated, cells were subjected to nuclear and cytosol fractionation and immunoblotted with antibodies against TFEB, PCNA, and β-actin. PCNA and β-actin were used as nuclear and cytosolic markers, respectively. ( c ) Immunoblot analysis of LC3 and SQSTM1/p62. ( d ) Immunoblot analysis of PEX5, p-70S6K, p-S6R, and p-4E-BP-1

Journal: Experimental & Molecular Medicine

Article Title: PEX5 regulates autophagy via the mTORC1-TFEB axis during starvation

doi: 10.1038/s12276-017-0007-8

Figure Lengend Snippet: Cells transfected with CTL siRNA (100 nM) and PEX5 siRNA (100 nM) for 24 h were pretreated with DMSO or rapamycin (1 μM) for 6 h, and then incubated in the absence of serum for 24 h. ( a ) After HepG2 cells were treated as indicated, cells were stained with an antibody against TFEB and analyzed by fluorescence microscopy. Scale bar: 10 μm. Data are presented as mean ± SD from three independent experiments (** P < 0.01). ( b ) After HepG2 cells were treated as indicated, cells were subjected to nuclear and cytosol fractionation and immunoblotted with antibodies against TFEB, PCNA, and β-actin. PCNA and β-actin were used as nuclear and cytosolic markers, respectively. ( c ) Immunoblot analysis of LC3 and SQSTM1/p62. ( d ) Immunoblot analysis of PEX5, p-70S6K, p-S6R, and p-4E-BP-1

Article Snippet: They were then transfected with 100 nM control siRNA, PEX5 siRNA (#1 sense-CGUAUCCUGGGAUCUCUCU, antisense-AGAGAGAUCCCAGGAUACG, #2 sense-GAAUUCAUCUCUGAAGUUA, antisense-UAACUUCAGAGAGAUGAAUUC-3, Bioneer, Daejeon, Korea), or TFEB siRNA (sc38509, Santa Cruz Biotechnology) for 24 h in serum-free medium, using Lipofectamine RNAiMax transfection reagent (Invitrogen/Life Technologies), and then exposed to serum starvation for 24 h. The effect of gene silencing was determined by Q-PCR and western blot analysis.

Techniques: Transfection, Incubation, Staining, Fluorescence, Microscopy, Fractionation, Western Blot

Cells transfected with siRNA for 24 h were pretreated with DMSO or rapamycin (1 μM) for 6 h, and then incubated in the absence of serum for 24 h. a Transcript levels of peroxisome-related genes were measured by Q-PCR. Values are expressed as fold increase compared to the starvation control group. Data are presented as mean ± s.d. from three independent experiments (** P < 0.01, * P < 0.05). b Immunoblot analysis of PMP70, ACOX1, DBP, catalase, and PEX5. c Q-PCR analysis of TFEB-regulated genes. Data are presented as mean ± s.d. from three independent experiments (** P < 0.01, * P < 0.05). d Protein expression of LAMP-1/LAMP-2 (lysosomal proteins) and cathepsins B/D (lysosomal proteases) were measured by immunoblotting. e Cells were transfected with empty, GFP-WT-TFEB, and GFP-TFEB-S142A plasmids. After 6 h of transfection, cells were treated with CTL and PEX5 siRNAs. After incubation in the absence of serum for 24 h, cells were subjected to immunoblot analysis with antibodies against PEX5, LC3, LAMP1, PMP70, TFEB, and β-actin

Journal: Experimental & Molecular Medicine

Article Title: PEX5 regulates autophagy via the mTORC1-TFEB axis during starvation

doi: 10.1038/s12276-017-0007-8

Figure Lengend Snippet: Cells transfected with siRNA for 24 h were pretreated with DMSO or rapamycin (1 μM) for 6 h, and then incubated in the absence of serum for 24 h. a Transcript levels of peroxisome-related genes were measured by Q-PCR. Values are expressed as fold increase compared to the starvation control group. Data are presented as mean ± s.d. from three independent experiments (** P < 0.01, * P < 0.05). b Immunoblot analysis of PMP70, ACOX1, DBP, catalase, and PEX5. c Q-PCR analysis of TFEB-regulated genes. Data are presented as mean ± s.d. from three independent experiments (** P < 0.01, * P < 0.05). d Protein expression of LAMP-1/LAMP-2 (lysosomal proteins) and cathepsins B/D (lysosomal proteases) were measured by immunoblotting. e Cells were transfected with empty, GFP-WT-TFEB, and GFP-TFEB-S142A plasmids. After 6 h of transfection, cells were treated with CTL and PEX5 siRNAs. After incubation in the absence of serum for 24 h, cells were subjected to immunoblot analysis with antibodies against PEX5, LC3, LAMP1, PMP70, TFEB, and β-actin

Article Snippet: They were then transfected with 100 nM control siRNA, PEX5 siRNA (#1 sense-CGUAUCCUGGGAUCUCUCU, antisense-AGAGAGAUCCCAGGAUACG, #2 sense-GAAUUCAUCUCUGAAGUUA, antisense-UAACUUCAGAGAGAUGAAUUC-3, Bioneer, Daejeon, Korea), or TFEB siRNA (sc38509, Santa Cruz Biotechnology) for 24 h in serum-free medium, using Lipofectamine RNAiMax transfection reagent (Invitrogen/Life Technologies), and then exposed to serum starvation for 24 h. The effect of gene silencing was determined by Q-PCR and western blot analysis.

Techniques: Transfection, Incubation, Control, Western Blot, Expressing