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control shrna  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology control shrna
    Control Shrna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1301 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/control+shrna/Control+shRNA+Lentiviral+Particles-A/pm42893040-52-6-16
    Average 96 stars, based on 1301 article reviews
    control shrna - by Bioz Stars, 2026-11
    96/100 stars

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    Related Articles

    Knockdown:

    Article Title: BIT1 as an Effector of EGFR-TKI-induced Apoptosis via TLE1 Inhibition in Lung Adenocarcinoma Cells.
    Article Snippet: .. To create stable knockdown cells lines, parental cell lines were transfected with 0.5 μg of control shRNA (cat. no.: sc-108080, Santa Cruz Biotechnology, Dallas, TX, USA) or TLE1-specific shRNA (cat. no.: sc-38558-V, Santa Cruz Biotechnology). .. After 48 h, transfected cells were treated with 1 μg/ml puromycin (cat. no.: A11138-02, Gibco) to select for stable clones.

    Transfection:

    Article Title: BIT1 as an Effector of EGFR-TKI-induced Apoptosis via TLE1 Inhibition in Lung Adenocarcinoma Cells.
    Article Snippet: .. To create stable knockdown cells lines, parental cell lines were transfected with 0.5 μg of control shRNA (cat. no.: sc-108080, Santa Cruz Biotechnology, Dallas, TX, USA) or TLE1-specific shRNA (cat. no.: sc-38558-V, Santa Cruz Biotechnology). .. After 48 h, transfected cells were treated with 1 μg/ml puromycin (cat. no.: A11138-02, Gibco) to select for stable clones.

    Article Title: FTO-Mediated m6A Demethylation of MZF1 Regulates DECR1 to Promote Fatty Acid Oxidation and Exacerbate Myocardial Ischemia/Reperfusion Injury : FTO-Mediated m6A Demethylation of MZF1 Enhances Fatty Acid Oxidation and Aggravates Myocardial I/R Injury.
    Article Snippet: Myocardial ischemia-reperfusion injury (MIRI) is a major clinical challenge, marked by metabolic disruptions and cellular damage following the restoration of blood flow after ischemia.. During ischemia, the heart shifts from fatty acid metabolism to glucose metabolism, leading to metabolic abnormalities, including reduced intracellular pH, ion disturbances, cell swelling, and apoptosis.. Upon reperfusion, fatty acid β-oxidation resumes, becoming the dominant energy source, which induces excessive oxidative stress and aggravates myocardial injury.

    Control:

    Article Title: BIT1 as an Effector of EGFR-TKI-induced Apoptosis via TLE1 Inhibition in Lung Adenocarcinoma Cells.
    Article Snippet: .. To create stable knockdown cells lines, parental cell lines were transfected with 0.5 μg of control shRNA (cat. no.: sc-108080, Santa Cruz Biotechnology, Dallas, TX, USA) or TLE1-specific shRNA (cat. no.: sc-38558-V, Santa Cruz Biotechnology). .. After 48 h, transfected cells were treated with 1 μg/ml puromycin (cat. no.: A11138-02, Gibco) to select for stable clones.

    Article Title: Context-Dependent Associations of HO-1 Modulation with Ferroptosis-Associated Injury and Vascular Cell Functions In Vitro.
    Article Snippet: HUVEC cells were seeded one day before transduction, after which 5 μg/mL polybrene (Sigma) and the lentiviral transduction particles were added. .. Control cells were transduced with control shRNA encoding a scrambled sequence using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    Article Title: FTO-Mediated m6A Demethylation of MZF1 Regulates DECR1 to Promote Fatty Acid Oxidation and Exacerbate Myocardial Ischemia/Reperfusion Injury : FTO-Mediated m6A Demethylation of MZF1 Enhances Fatty Acid Oxidation and Aggravates Myocardial I/R Injury.
    Article Snippet: Myocardial ischemia-reperfusion injury (MIRI) is a major clinical challenge, marked by metabolic disruptions and cellular damage following the restoration of blood flow after ischemia.. During ischemia, the heart shifts from fatty acid metabolism to glucose metabolism, leading to metabolic abnormalities, including reduced intracellular pH, ion disturbances, cell swelling, and apoptosis.. Upon reperfusion, fatty acid β-oxidation resumes, becoming the dominant energy source, which induces excessive oxidative stress and aggravates myocardial injury.

    Article Title: Canonical BAF chromatin remodeling complex specifies stem cell fate via cell-type-specific co-factor recruitment.
    Article Snippet: For lentiviral injection, a 10 μl Hamilton syringe (Hamilton Company, Reno, NV) connected to a glass capillary micropipette was inserted into the bone defect at the proximal end of the incisor. .. Subsequently, 3 μl of lentivirus carrying Runx2 shRNA (Santa Cruz, sc-37146-v) or control shRNA (Santa Cruz, sc-108080) was injected into the proximal end of the incisor mesenchyme at a flow rate of 20 nl/s. ..

    Article Title: Dysregulation of a novel autophagosome-mitochondria contact contributes to autophagy dysfunction and neurodegeneration in tauopathy
    Article Snippet: .. Sources of antibodies or reagents are as follows: polyclonal antibodies against LC3 (Cell Signaling Technology, Cat# 2775), AMPKα (Cell Signaling Technology, Cat# 2532), p62/SQSTM1 (Abnova, Cat# H00008878-M01), NeuN (Millipore/Sigma, Cat# ABN78), TBC1D15 (Abcam, Cat# ab121396), Tau5 (DAKO, Cat# A0024), and synaptophysin/SYP (Abcam, Cat# ab32127); monoclonal antibodies against phospho-AMPKα (Cell Signaling Technology, Cat# 2535), AT8 (ThermoFisher Scientific, Cat# MN1020), syntaxin 1/STX1 (Santa Cruz Biotechnology, Cat# sc-12736), Rab7 (Sigma, Cat# R8779), GAPDH (Sigma, Cat# CB1001), TOM20 (Abcam, Cat# ab186734), mCherry (Takara, Cat# 632543), and Alexa fluor 488- (Cat# A-11017; Cat# A-11070), 546- (Cat# A-11018; Cat# A-11071), and 647- (Cat# A-21237; Cat# A-21246) conjugated secondary antibodies (Invitrogen); 10-NCP (VWR, Cat# 80017-188), trehalose (Cat# T0167), cycloheximide (Cat# 01810), rotenone (Cat# 557368), antimycin A (Cat# 1397-94-0), AICAR (Cat# A9978), CC (Cat# 171264), epoxomicin (Cat# 324800), CID 1067700 (Cat# SML0545), DMSO (Cat# D2650), and Sarkosyl (Cat# L9150) (Sigma); TBC1D15 shRNA (Cat# sc-154093-SH), Fis1 shRNA (Cat# sc-60644-SH), and control shRNA (Cat# sc-108060) (Santa Cruz Biotechnology); MitoView TM Green (Cat# 70054) (biotium); LentiBrite RFP-LC3 Lentiviral Biosensor (Cat# 17-10143) (Sigma). .. The constructs encoding GFP-LC3, DsRed-Mito, and mRFP-LC3 were prepared as we previously described ( , , , ). mCherry-TBC1D15 and mCherry-TBC1D15 D397A were from Vector Biolabs.

    Article Title: SMARCA4 deficiency in glioblastoma: Mitochondrial transfer from MSCs and the clinical dilemma in targeting the tumor microenvironment
    Article Snippet: .. Human MSCs were transduced with lentiviral particles carrying either SMARCA4 shRNA (sc-29827-V, Santa Cruz) or scrambled control shRNA (sc-108080, Santa Cruz). ..

    Article Title: Metformin and caffeic acid phenethyl ester counteract hyperglycemia-induced endothelial dysfunction through growth differentiation factor 15 and heme oxygenase-1-associated signaling.
    Article Snippet: Following this, 5 μg/mL of polybrene (Sigma-Aldrich Co.) and the lentiviral particles were introduced. .. Control cells were transduced with a control shRNA encoding a scrambled sequence, using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    shRNA:

    Article Title: BIT1 as an Effector of EGFR-TKI-induced Apoptosis via TLE1 Inhibition in Lung Adenocarcinoma Cells.
    Article Snippet: .. To create stable knockdown cells lines, parental cell lines were transfected with 0.5 μg of control shRNA (cat. no.: sc-108080, Santa Cruz Biotechnology, Dallas, TX, USA) or TLE1-specific shRNA (cat. no.: sc-38558-V, Santa Cruz Biotechnology). .. After 48 h, transfected cells were treated with 1 μg/ml puromycin (cat. no.: A11138-02, Gibco) to select for stable clones.

    Article Title: Context-Dependent Associations of HO-1 Modulation with Ferroptosis-Associated Injury and Vascular Cell Functions In Vitro.
    Article Snippet: HUVEC cells were seeded one day before transduction, after which 5 μg/mL polybrene (Sigma) and the lentiviral transduction particles were added. .. Control cells were transduced with control shRNA encoding a scrambled sequence using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    Article Title: FTO-Mediated m6A Demethylation of MZF1 Regulates DECR1 to Promote Fatty Acid Oxidation and Exacerbate Myocardial Ischemia/Reperfusion Injury : FTO-Mediated m6A Demethylation of MZF1 Enhances Fatty Acid Oxidation and Aggravates Myocardial I/R Injury.
    Article Snippet: Myocardial ischemia-reperfusion injury (MIRI) is a major clinical challenge, marked by metabolic disruptions and cellular damage following the restoration of blood flow after ischemia.. During ischemia, the heart shifts from fatty acid metabolism to glucose metabolism, leading to metabolic abnormalities, including reduced intracellular pH, ion disturbances, cell swelling, and apoptosis.. Upon reperfusion, fatty acid β-oxidation resumes, becoming the dominant energy source, which induces excessive oxidative stress and aggravates myocardial injury.

    Article Title: Canonical BAF chromatin remodeling complex specifies stem cell fate via cell-type-specific co-factor recruitment.
    Article Snippet: For lentiviral injection, a 10 μl Hamilton syringe (Hamilton Company, Reno, NV) connected to a glass capillary micropipette was inserted into the bone defect at the proximal end of the incisor. .. Subsequently, 3 μl of lentivirus carrying Runx2 shRNA (Santa Cruz, sc-37146-v) or control shRNA (Santa Cruz, sc-108080) was injected into the proximal end of the incisor mesenchyme at a flow rate of 20 nl/s. ..

    Article Title: Dysregulation of a novel autophagosome-mitochondria contact contributes to autophagy dysfunction and neurodegeneration in tauopathy
    Article Snippet: .. Sources of antibodies or reagents are as follows: polyclonal antibodies against LC3 (Cell Signaling Technology, Cat# 2775), AMPKα (Cell Signaling Technology, Cat# 2532), p62/SQSTM1 (Abnova, Cat# H00008878-M01), NeuN (Millipore/Sigma, Cat# ABN78), TBC1D15 (Abcam, Cat# ab121396), Tau5 (DAKO, Cat# A0024), and synaptophysin/SYP (Abcam, Cat# ab32127); monoclonal antibodies against phospho-AMPKα (Cell Signaling Technology, Cat# 2535), AT8 (ThermoFisher Scientific, Cat# MN1020), syntaxin 1/STX1 (Santa Cruz Biotechnology, Cat# sc-12736), Rab7 (Sigma, Cat# R8779), GAPDH (Sigma, Cat# CB1001), TOM20 (Abcam, Cat# ab186734), mCherry (Takara, Cat# 632543), and Alexa fluor 488- (Cat# A-11017; Cat# A-11070), 546- (Cat# A-11018; Cat# A-11071), and 647- (Cat# A-21237; Cat# A-21246) conjugated secondary antibodies (Invitrogen); 10-NCP (VWR, Cat# 80017-188), trehalose (Cat# T0167), cycloheximide (Cat# 01810), rotenone (Cat# 557368), antimycin A (Cat# 1397-94-0), AICAR (Cat# A9978), CC (Cat# 171264), epoxomicin (Cat# 324800), CID 1067700 (Cat# SML0545), DMSO (Cat# D2650), and Sarkosyl (Cat# L9150) (Sigma); TBC1D15 shRNA (Cat# sc-154093-SH), Fis1 shRNA (Cat# sc-60644-SH), and control shRNA (Cat# sc-108060) (Santa Cruz Biotechnology); MitoView TM Green (Cat# 70054) (biotium); LentiBrite RFP-LC3 Lentiviral Biosensor (Cat# 17-10143) (Sigma). .. The constructs encoding GFP-LC3, DsRed-Mito, and mRFP-LC3 were prepared as we previously described ( , , , ). mCherry-TBC1D15 and mCherry-TBC1D15 D397A were from Vector Biolabs.

    Article Title: SMARCA4 deficiency in glioblastoma: Mitochondrial transfer from MSCs and the clinical dilemma in targeting the tumor microenvironment
    Article Snippet: .. Human MSCs were transduced with lentiviral particles carrying either SMARCA4 shRNA (sc-29827-V, Santa Cruz) or scrambled control shRNA (sc-108080, Santa Cruz). ..

    Article Title: Metformin and caffeic acid phenethyl ester counteract hyperglycemia-induced endothelial dysfunction through growth differentiation factor 15 and heme oxygenase-1-associated signaling.
    Article Snippet: Following this, 5 μg/mL of polybrene (Sigma-Aldrich Co.) and the lentiviral particles were introduced. .. Control cells were transduced with a control shRNA encoding a scrambled sequence, using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    Transduction:

    Article Title: Context-Dependent Associations of HO-1 Modulation with Ferroptosis-Associated Injury and Vascular Cell Functions In Vitro.
    Article Snippet: HUVEC cells were seeded one day before transduction, after which 5 μg/mL polybrene (Sigma) and the lentiviral transduction particles were added. .. Control cells were transduced with control shRNA encoding a scrambled sequence using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    Article Title: SMARCA4 deficiency in glioblastoma: Mitochondrial transfer from MSCs and the clinical dilemma in targeting the tumor microenvironment
    Article Snippet: .. Human MSCs were transduced with lentiviral particles carrying either SMARCA4 shRNA (sc-29827-V, Santa Cruz) or scrambled control shRNA (sc-108080, Santa Cruz). ..

    Article Title: SMARCA4 deficiency in glioblastoma: Mitochondrial transfer from MSCs and the clinical dilemma in targeting the tumor microenvironment.
    Article Snippet: .. Human MSCs were transduced with lentiviral particles carrying either SMARCA4 shRNA (sc-29827-V, Santa Cruz) or scrambled control shRNA (sc-108080, Santa Cruz). ..

    Article Title: Metformin and caffeic acid phenethyl ester counteract hyperglycemia-induced endothelial dysfunction through growth differentiation factor 15 and heme oxygenase-1-associated signaling.
    Article Snippet: Following this, 5 μg/mL of polybrene (Sigma-Aldrich Co.) and the lentiviral particles were introduced. .. Control cells were transduced with a control shRNA encoding a scrambled sequence, using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    Sequencing:

    Article Title: Context-Dependent Associations of HO-1 Modulation with Ferroptosis-Associated Injury and Vascular Cell Functions In Vitro.
    Article Snippet: HUVEC cells were seeded one day before transduction, after which 5 μg/mL polybrene (Sigma) and the lentiviral transduction particles were added. .. Control cells were transduced with control shRNA encoding a scrambled sequence using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    Article Title: Metformin and caffeic acid phenethyl ester counteract hyperglycemia-induced endothelial dysfunction through growth differentiation factor 15 and heme oxygenase-1-associated signaling.
    Article Snippet: Following this, 5 μg/mL of polybrene (Sigma-Aldrich Co.) and the lentiviral particles were introduced. .. Control cells were transduced with a control shRNA encoding a scrambled sequence, using transduction particles (sc-108080, Santa Cruz Biotechnology). ..

    Injection:

    Article Title: Canonical BAF chromatin remodeling complex specifies stem cell fate via cell-type-specific co-factor recruitment.
    Article Snippet: For lentiviral injection, a 10 μl Hamilton syringe (Hamilton Company, Reno, NV) connected to a glass capillary micropipette was inserted into the bone defect at the proximal end of the incisor. .. Subsequently, 3 μl of lentivirus carrying Runx2 shRNA (Santa Cruz, sc-37146-v) or control shRNA (Santa Cruz, sc-108080) was injected into the proximal end of the incisor mesenchyme at a flow rate of 20 nl/s. ..

    Bioprocessing:

    Article Title: Dysregulation of a novel autophagosome-mitochondria contact contributes to autophagy dysfunction and neurodegeneration in tauopathy
    Article Snippet: .. Sources of antibodies or reagents are as follows: polyclonal antibodies against LC3 (Cell Signaling Technology, Cat# 2775), AMPKα (Cell Signaling Technology, Cat# 2532), p62/SQSTM1 (Abnova, Cat# H00008878-M01), NeuN (Millipore/Sigma, Cat# ABN78), TBC1D15 (Abcam, Cat# ab121396), Tau5 (DAKO, Cat# A0024), and synaptophysin/SYP (Abcam, Cat# ab32127); monoclonal antibodies against phospho-AMPKα (Cell Signaling Technology, Cat# 2535), AT8 (ThermoFisher Scientific, Cat# MN1020), syntaxin 1/STX1 (Santa Cruz Biotechnology, Cat# sc-12736), Rab7 (Sigma, Cat# R8779), GAPDH (Sigma, Cat# CB1001), TOM20 (Abcam, Cat# ab186734), mCherry (Takara, Cat# 632543), and Alexa fluor 488- (Cat# A-11017; Cat# A-11070), 546- (Cat# A-11018; Cat# A-11071), and 647- (Cat# A-21237; Cat# A-21246) conjugated secondary antibodies (Invitrogen); 10-NCP (VWR, Cat# 80017-188), trehalose (Cat# T0167), cycloheximide (Cat# 01810), rotenone (Cat# 557368), antimycin A (Cat# 1397-94-0), AICAR (Cat# A9978), CC (Cat# 171264), epoxomicin (Cat# 324800), CID 1067700 (Cat# SML0545), DMSO (Cat# D2650), and Sarkosyl (Cat# L9150) (Sigma); TBC1D15 shRNA (Cat# sc-154093-SH), Fis1 shRNA (Cat# sc-60644-SH), and control shRNA (Cat# sc-108060) (Santa Cruz Biotechnology); MitoView TM Green (Cat# 70054) (biotium); LentiBrite RFP-LC3 Lentiviral Biosensor (Cat# 17-10143) (Sigma). .. The constructs encoding GFP-LC3, DsRed-Mito, and mRFP-LC3 were prepared as we previously described ( , , , ). mCherry-TBC1D15 and mCherry-TBC1D15 D397A were from Vector Biolabs.



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    Effect of Pz modulation on ASM proliferation and Wnt signaling. The effect of Pz1 agonist (Yoda1) and antagonist (GsMTx4) on ASM cell proliferation was evaluated using the CyQuant assay in all 4 groups (A). Baseline cell proliferation for young non-asthmatic ASM is indicated by the blue dotted line and used for comparison. Cells were treated with vehicle, Yoda1 (10 µm), and GsMTx4 (1 µm). No consistent pattern of drug, age, disease, or Pz modulation was noted on ASM cell proliferation. Expression of canonical Wnt (Wnt3a) (B), and non-canonical Wnt5ab (C) β-catenin (D) and GSK3β (E) elements was determined in cells where Pz1 channel expression was suppressed by <t>shRNA.</t> ASM cells failed to show the expected increase in these elements, even when exposed to mechanical stretch (representative electropherograms are shown in (F). Cells were verified for knockdown efficiency by using standard Western blot analysis (G) ** P < .01 vs negative shRNA. N = 5-6 patients per group, mean ± SEM for box plots.
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    Relationship between MIF and TMED10. (A) Secretion of MIF was measured in HEK293T cells either untreated or treated with brefeldin A. (B) Secretion of MIF was measured in HEK293T cells untreated or treated with the indicated doses of probenecid. (C) GST or the indicated GST-TMED10 domains coupled to glutathione sepharose beads were incubated with whole cell lysates from HEK293T cells expressing Flag-MIF. Elutes were subjected to SDS-PAGE and immunoblotted with an anti-Flag antibody. (D) GST-MIF coupled to glutathione sepharose beads was incubated with whole cell lysate from HEK293T cells expressing GFP-TMED10 or GFP-TMED10(ΔCT). Elutes were subjected to SDS-PAGE and immunoblotted with anti-GFP antibody. (E) Secretion of MIF was measured in HEK293T cells expressing mock or GFP-TMED10. (F) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting TMED10 for 72 h. (G) A crosslink assay was performed in HEK293T cells expressing GFP-TMED10 with mock or Flag-MIF to examine the oligomeric form of TMED10. (H) Secretion of Flag-MIF was measured in HEK293T cells expressing mock, RAB1A, 1B, and 2A.

    Journal: Autophagy Reports

    Article Title: TRPML3 and TRPML1/3 differentially regulate unconventional MIF secretion through TMED10 and secretory autophagy

    doi: 10.1080/27694127.2026.2740425

    Figure Lengend Snippet: Relationship between MIF and TMED10. (A) Secretion of MIF was measured in HEK293T cells either untreated or treated with brefeldin A. (B) Secretion of MIF was measured in HEK293T cells untreated or treated with the indicated doses of probenecid. (C) GST or the indicated GST-TMED10 domains coupled to glutathione sepharose beads were incubated with whole cell lysates from HEK293T cells expressing Flag-MIF. Elutes were subjected to SDS-PAGE and immunoblotted with an anti-Flag antibody. (D) GST-MIF coupled to glutathione sepharose beads was incubated with whole cell lysate from HEK293T cells expressing GFP-TMED10 or GFP-TMED10(ΔCT). Elutes were subjected to SDS-PAGE and immunoblotted with anti-GFP antibody. (E) Secretion of MIF was measured in HEK293T cells expressing mock or GFP-TMED10. (F) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting TMED10 for 72 h. (G) A crosslink assay was performed in HEK293T cells expressing GFP-TMED10 with mock or Flag-MIF to examine the oligomeric form of TMED10. (H) Secretion of Flag-MIF was measured in HEK293T cells expressing mock, RAB1A, 1B, and 2A.

    Article Snippet: For knockdown experiments, siRNA targeting human TMED10 (BIONEER, 10,972–1), human ATG5 (BIONEER, 9474–1), human SYT5 (BIONEER, 6861–1), and a negative control scrambled siRNA (Santa Cruz Biotechnology, sc-37007) were used.

    Techniques: Incubation, Expressing, SDS Page, Control

    Effect of autophagy on MIF secretion. (A) HeLa cells expressing GFP-LC3 and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy. (B) Secretion of MIF was measured in HEK293T cells either untreated or treated with 10 μM SAR405. (C) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting ATG5 for 72 h. (D) Secretion of MIF was measured in HEK293T cells either untreated or treated with 100 nM bafilomycin A1. (E) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting SYT5 for 48 h. Knockdown efficiency was confirmed by immunoblotting for SYT5. (F) Secretion of Flag-MIF was measured in HEK293T cells expressing mock or GFP-SEC22B. (G) HeLa cells expressing GFP-SEC22B and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy. (H, upper) HeLa cells co-expressing GFP-MIF and Flag-VAMP4 were treated with cold digitonin (50 µg/ml), immunostained with an anti-Flag antibody, and imaged by fluorescence microscopy. (H, lower) HeLa cells expressing GFP-VAMP8 and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy.

    Journal: Autophagy Reports

    Article Title: TRPML3 and TRPML1/3 differentially regulate unconventional MIF secretion through TMED10 and secretory autophagy

    doi: 10.1080/27694127.2026.2740425

    Figure Lengend Snippet: Effect of autophagy on MIF secretion. (A) HeLa cells expressing GFP-LC3 and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy. (B) Secretion of MIF was measured in HEK293T cells either untreated or treated with 10 μM SAR405. (C) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting ATG5 for 72 h. (D) Secretion of MIF was measured in HEK293T cells either untreated or treated with 100 nM bafilomycin A1. (E) Secretion of MIF was measured in HEK293T cells treated with control siRNA or siRNA targeting SYT5 for 48 h. Knockdown efficiency was confirmed by immunoblotting for SYT5. (F) Secretion of Flag-MIF was measured in HEK293T cells expressing mock or GFP-SEC22B. (G) HeLa cells expressing GFP-SEC22B and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy. (H, upper) HeLa cells co-expressing GFP-MIF and Flag-VAMP4 were treated with cold digitonin (50 µg/ml), immunostained with an anti-Flag antibody, and imaged by fluorescence microscopy. (H, lower) HeLa cells expressing GFP-VAMP8 and mCherry-MIF were treated with cold digitonin (50 µg/ml) and imaged by fluorescence microscopy.

    Article Snippet: For knockdown experiments, siRNA targeting human TMED10 (BIONEER, 10,972–1), human ATG5 (BIONEER, 9474–1), human SYT5 (BIONEER, 6861–1), and a negative control scrambled siRNA (Santa Cruz Biotechnology, sc-37007) were used.

    Techniques: Expressing, Fluorescence, Microscopy, Control, Knockdown, Western Blot

    Effect of Pz modulation on ASM proliferation and Wnt signaling. The effect of Pz1 agonist (Yoda1) and antagonist (GsMTx4) on ASM cell proliferation was evaluated using the CyQuant assay in all 4 groups (A). Baseline cell proliferation for young non-asthmatic ASM is indicated by the blue dotted line and used for comparison. Cells were treated with vehicle, Yoda1 (10 µm), and GsMTx4 (1 µm). No consistent pattern of drug, age, disease, or Pz modulation was noted on ASM cell proliferation. Expression of canonical Wnt (Wnt3a) (B), and non-canonical Wnt5ab (C) β-catenin (D) and GSK3β (E) elements was determined in cells where Pz1 channel expression was suppressed by shRNA. ASM cells failed to show the expected increase in these elements, even when exposed to mechanical stretch (representative electropherograms are shown in (F). Cells were verified for knockdown efficiency by using standard Western blot analysis (G) ** P < .01 vs negative shRNA. N = 5-6 patients per group, mean ± SEM for box plots.

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: Piezo1 mediates stretch-induced Wnt/β-catenin signaling and extracellular matrix remodeling in aging asthmatic airway smooth muscle

    doi: 10.1093/ajrcmb/aanag058

    Figure Lengend Snippet: Effect of Pz modulation on ASM proliferation and Wnt signaling. The effect of Pz1 agonist (Yoda1) and antagonist (GsMTx4) on ASM cell proliferation was evaluated using the CyQuant assay in all 4 groups (A). Baseline cell proliferation for young non-asthmatic ASM is indicated by the blue dotted line and used for comparison. Cells were treated with vehicle, Yoda1 (10 µm), and GsMTx4 (1 µm). No consistent pattern of drug, age, disease, or Pz modulation was noted on ASM cell proliferation. Expression of canonical Wnt (Wnt3a) (B), and non-canonical Wnt5ab (C) β-catenin (D) and GSK3β (E) elements was determined in cells where Pz1 channel expression was suppressed by shRNA. ASM cells failed to show the expected increase in these elements, even when exposed to mechanical stretch (representative electropherograms are shown in (F). Cells were verified for knockdown efficiency by using standard Western blot analysis (G) ** P < .01 vs negative shRNA. N = 5-6 patients per group, mean ± SEM for box plots.

    Article Snippet: Control shRNA lentiviral particles-A (sc-108080) and Pz1 shRNA lentiviral particles (sc-93227-V) were procured from Santa Cruz Biotechnology (TX, USA).

    Techniques: CyQUANT Assay, Comparison, Expressing, shRNA, Knockdown, Western Blot, Analysis