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sequence shrna  (Santa Cruz Biotechnology)


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    Santa Cruz Biotechnology sequence shrna
    a Schematic <t>of</t> <t>Gck</t> virogenetic <t>shRNA</t> or control shRNA silencing surgery in the major taste fields of the tongue. b Schematic of brief access taste test of maltose and sucrose in lickometer. c Mean (±SEM) lick score of 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=11) and GCK KD (n=12) TRPM5+. d Mean lick scores averaged across concentration for control and GCK KD TRPM5+ (11-12/group). e Mean (±SEM) 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=10) and GCK KD (n=11) TRPM5-. f Mean lick scores averaged across concentration for control and GCK KD TRPM5- (10-11/group). g Mean relative transcript expression of Mgam for B6 (n=8), TRPM5+ (n=8) and TRPM5- (n=9). h Mean (±SEM) relative MGAM transcript for B6 sugar naïve (n=2), glucose experience (n=5), fructose experience (n=6), or glucose + fructose experience(n=5). (*:p<0.05, **:p<0.01,***:p<0.001,****:p<0.0001). All tests were conducted in the Davis Rig. Statistical Analysis are in Supplementary Table 3.
    Sequence Shrna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1290 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/control+sirna+lentiviral+particles/Control+shRNA+Lentiviral+Particles-A/bio_rxiv__64898__2026__03__13__710876-267-18-22
    Average 96 stars, based on 1290 article reviews
    sequence shrna - by Bioz Stars, 2026-09
    96/100 stars

    Images

    1) Product Images from "An enzymatic–metabolic sensing axis in taste cells detects glucose-yielding carbohydrates"

    Article Title: An enzymatic–metabolic sensing axis in taste cells detects glucose-yielding carbohydrates

    Journal: bioRxiv

    doi: 10.64898/2026.03.13.710876

    a Schematic of Gck virogenetic shRNA or control shRNA silencing surgery in the major taste fields of the tongue. b Schematic of brief access taste test of maltose and sucrose in lickometer. c Mean (±SEM) lick score of 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=11) and GCK KD (n=12) TRPM5+. d Mean lick scores averaged across concentration for control and GCK KD TRPM5+ (11-12/group). e Mean (±SEM) 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=10) and GCK KD (n=11) TRPM5-. f Mean lick scores averaged across concentration for control and GCK KD TRPM5- (10-11/group). g Mean relative transcript expression of Mgam for B6 (n=8), TRPM5+ (n=8) and TRPM5- (n=9). h Mean (±SEM) relative MGAM transcript for B6 sugar naïve (n=2), glucose experience (n=5), fructose experience (n=6), or glucose + fructose experience(n=5). (*:p<0.05, **:p<0.01,***:p<0.001,****:p<0.0001). All tests were conducted in the Davis Rig. Statistical Analysis are in Supplementary Table 3.
    Figure Legend Snippet: a Schematic of Gck virogenetic shRNA or control shRNA silencing surgery in the major taste fields of the tongue. b Schematic of brief access taste test of maltose and sucrose in lickometer. c Mean (±SEM) lick score of 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=11) and GCK KD (n=12) TRPM5+. d Mean lick scores averaged across concentration for control and GCK KD TRPM5+ (11-12/group). e Mean (±SEM) 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=10) and GCK KD (n=11) TRPM5-. f Mean lick scores averaged across concentration for control and GCK KD TRPM5- (10-11/group). g Mean relative transcript expression of Mgam for B6 (n=8), TRPM5+ (n=8) and TRPM5- (n=9). h Mean (±SEM) relative MGAM transcript for B6 sugar naïve (n=2), glucose experience (n=5), fructose experience (n=6), or glucose + fructose experience(n=5). (*:p<0.05, **:p<0.01,***:p<0.001,****:p<0.0001). All tests were conducted in the Davis Rig. Statistical Analysis are in Supplementary Table 3.

    Techniques Used: shRNA, Control, Concentration Assay, Expressing

    a Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Mean relative transcript expression of Mgam, Gck, and Tas1r3 in control (n= 4) and MGAM KD (n=4) in TRPM5+ mice c Schematic of burst structure (period of continuous licks separated by <1s) and time frame d Schematic of 300 lick test of 0.6M maltose in lickometer. e Schematical graph showing relationship between burst size and hedonic value. f Mean (±SEM) burst size, burst number, total licks for 300 lick test in control (n=3) and MGAM KD (n=3) in TRPM5+ mice (*:p<0.05). All tests were conducted in the gustometer. Statistical Analysis are in .
    Figure Legend Snippet: a Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Mean relative transcript expression of Mgam, Gck, and Tas1r3 in control (n= 4) and MGAM KD (n=4) in TRPM5+ mice c Schematic of burst structure (period of continuous licks separated by <1s) and time frame d Schematic of 300 lick test of 0.6M maltose in lickometer. e Schematical graph showing relationship between burst size and hedonic value. f Mean (±SEM) burst size, burst number, total licks for 300 lick test in control (n=3) and MGAM KD (n=3) in TRPM5+ mice (*:p<0.05). All tests were conducted in the gustometer. Statistical Analysis are in .

    Techniques Used: Control, shRNA, Expressing

    a Schematic depicting T1R2+T1R3 sweet receptor transgenic knockout mouse. b Mean transcript expression of Gck and Mgam for sugar naive (n=4) and sugar exposed (n=4) T1RKO. c Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. d Schematic of brief access taste test of maltose and sucrose in lickometer. e Block 1 mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=5) and MGAM KD (n=5) T1RKO. f Mean lick scores averaged across concentration for control and MGAM KD for first block (5/group). g Schematic of brief access taste test of glucose and fructose in lickometer. h First block mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M glucose and fructose in control (n=6) and MGAM KD (n=5) T1RKO. i First block glucose and fructose mean lick scores averaged across concentration for control (n=6) and MGAM KD (n=5) T1RKO. All tests were conducted in the Davis Rig. (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). Statistical Analysis are in Supplementary Table 7.
    Figure Legend Snippet: a Schematic depicting T1R2+T1R3 sweet receptor transgenic knockout mouse. b Mean transcript expression of Gck and Mgam for sugar naive (n=4) and sugar exposed (n=4) T1RKO. c Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. d Schematic of brief access taste test of maltose and sucrose in lickometer. e Block 1 mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=5) and MGAM KD (n=5) T1RKO. f Mean lick scores averaged across concentration for control and MGAM KD for first block (5/group). g Schematic of brief access taste test of glucose and fructose in lickometer. h First block mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M glucose and fructose in control (n=6) and MGAM KD (n=5) T1RKO. i First block glucose and fructose mean lick scores averaged across concentration for control (n=6) and MGAM KD (n=5) T1RKO. All tests were conducted in the Davis Rig. (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). Statistical Analysis are in Supplementary Table 7.

    Techniques Used: Transgenic Assay, Knock-Out, Expressing, Control, shRNA, Blocking Assay, Concentration Assay

    a Schematic showing control, Gck, and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Schematic of 1000 lick test of mixed nutrient solution (Peptamen) in lickometer for pre-surgery and post-surgery. c Mean total licks for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. d Mean meal duration for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. e Mean burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. f Mean burst number for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. g Mean (±SEM) cumulative first 15 burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. (n=12/group for all tests) (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). All tests were conducted in the gustometer. Statistical Analysis are in Supplementary Table 8.
    Figure Legend Snippet: a Schematic showing control, Gck, and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Schematic of 1000 lick test of mixed nutrient solution (Peptamen) in lickometer for pre-surgery and post-surgery. c Mean total licks for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. d Mean meal duration for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. e Mean burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. f Mean burst number for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. g Mean (±SEM) cumulative first 15 burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. (n=12/group for all tests) (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). All tests were conducted in the gustometer. Statistical Analysis are in Supplementary Table 8.

    Techniques Used: Control, shRNA

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    Incubation:

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    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
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    Transduction:

    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
    Article Snippet: Transfection and CARM1 gene silencing— To identify the effects of genetic knock-down of CARM1 on dendritic morphology and synaptic clustering of synapsin, NR2B, and PSD-95, hippocampal neurons were first transfected with the construct pCAG-mGFP-Actin [a gift from Ryohei Yasuda (Addgene plasmid # 21948)] at 7 DIV with Lipofectamine 2000 (Invitrogen) following the manufacturer’s protocol. .. After a 2day incubation, neurons were untreated or transduced with rat CARM1or PRMT1-specific siRNA or scrambled control siRNA lentiviral particles (5 x 10 3 viral particles/μl, Santa Cruz) at a multiplicity of infection (MOI) of 4 to ensure efficient infection following the manufacturer’s protocol, followed by overnight incubation. ..

    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
    Article Snippet: To identify the effects of genetic knockdown of CARM1 on dendritic morphology and synaptic clustering of synapsin, NR2B, and PSD-95, hippocampal neurons were first transfected with the construct pCAG-mGFP-actin (a gift from Ryohei Yasuda (Addgene plasmid number 21948)) at 7 DIV with Lipofectamine 2000 (Invitrogen) following the manufacturer's protocol. .. After a 2-day incubation, neurons were untreated or transduced with rat CARM1- or PRMT1-specific siRNA or scrambled control siRNA lentiviral particles (5 × 10 3 viral particles/μl) (Santa Cruz Biotechnology) at a multiplicity of infection (m.o.i.) of 4 to ensure efficient infection following the manufacturer's protocol, followed by overnight incubation. ..

    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
    Article Snippet: Transfection and CARM1 gene silencing— To identify the effects of genetic knock-down of CARM1 on dendritic morphology and synaptic clustering of synapsin, NR2B, and PSD-95, hippocampal neurons were first transfected with the construct pCAG-mGFP-Actin [a gift from Ryohei Yasuda (Addgene plasmid # 21948)] at 7 DIV with Lipofectamine 2000 (Invitrogen) following the manufacturer’s protocol. .. After a 2- day incubation, neurons were untreated or transduced with rat CARM1- or PRMT1-specific siRNA or scrambled control siRNA lentiviral particles (5 x 10 3 viral particles/μl, Santa Cruz) at a multiplicity of infection (MOI) of 4 to ensure efficient infection following the manufacturer’s protocol, followed by overnight incubation. ..

    Control:

    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
    Article Snippet: Transfection and CARM1 gene silencing— To identify the effects of genetic knock-down of CARM1 on dendritic morphology and synaptic clustering of synapsin, NR2B, and PSD-95, hippocampal neurons were first transfected with the construct pCAG-mGFP-Actin [a gift from Ryohei Yasuda (Addgene plasmid # 21948)] at 7 DIV with Lipofectamine 2000 (Invitrogen) following the manufacturer’s protocol. .. After a 2day incubation, neurons were untreated or transduced with rat CARM1or PRMT1-specific siRNA or scrambled control siRNA lentiviral particles (5 x 10 3 viral particles/μl, Santa Cruz) at a multiplicity of infection (MOI) of 4 to ensure efficient infection following the manufacturer’s protocol, followed by overnight incubation. ..

    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
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    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
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    Infection:

    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
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    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
    Article Snippet: To identify the effects of genetic knockdown of CARM1 on dendritic morphology and synaptic clustering of synapsin, NR2B, and PSD-95, hippocampal neurons were first transfected with the construct pCAG-mGFP-actin (a gift from Ryohei Yasuda (Addgene plasmid number 21948)) at 7 DIV with Lipofectamine 2000 (Invitrogen) following the manufacturer's protocol. .. After a 2-day incubation, neurons were untreated or transduced with rat CARM1- or PRMT1-specific siRNA or scrambled control siRNA lentiviral particles (5 × 10 3 viral particles/μl) (Santa Cruz Biotechnology) at a multiplicity of infection (m.o.i.) of 4 to ensure efficient infection following the manufacturer's protocol, followed by overnight incubation. ..

    Article Title: Inhibition of coactivator-associated arginine methyltransferase 1 modulates dendritic arborization and spine maturation of cultured hippocampal neurons
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    a Schematic of Gck virogenetic shRNA or control shRNA silencing surgery in the major taste fields of the tongue. b Schematic of brief access taste test of maltose and sucrose in lickometer. c Mean (±SEM) lick score of 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=11) and GCK KD (n=12) TRPM5+. d Mean lick scores averaged across concentration for control and GCK KD TRPM5+ (11-12/group). e Mean (±SEM) 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=10) and GCK KD (n=11) TRPM5-. f Mean lick scores averaged across concentration for control and GCK KD TRPM5- (10-11/group). g Mean relative transcript expression of Mgam for B6 (n=8), TRPM5+ (n=8) and TRPM5- (n=9). h Mean (±SEM) relative MGAM transcript for B6 sugar naïve (n=2), glucose experience (n=5), fructose experience (n=6), or glucose + fructose experience(n=5). (*:p<0.05, **:p<0.01,***:p<0.001,****:p<0.0001). All tests were conducted in the Davis Rig. Statistical Analysis are in Supplementary Table 3.

    Journal: bioRxiv

    Article Title: An enzymatic–metabolic sensing axis in taste cells detects glucose-yielding carbohydrates

    doi: 10.64898/2026.03.13.710876

    Figure Lengend Snippet: a Schematic of Gck virogenetic shRNA or control shRNA silencing surgery in the major taste fields of the tongue. b Schematic of brief access taste test of maltose and sucrose in lickometer. c Mean (±SEM) lick score of 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=11) and GCK KD (n=12) TRPM5+. d Mean lick scores averaged across concentration for control and GCK KD TRPM5+ (11-12/group). e Mean (±SEM) 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=10) and GCK KD (n=11) TRPM5-. f Mean lick scores averaged across concentration for control and GCK KD TRPM5- (10-11/group). g Mean relative transcript expression of Mgam for B6 (n=8), TRPM5+ (n=8) and TRPM5- (n=9). h Mean (±SEM) relative MGAM transcript for B6 sugar naïve (n=2), glucose experience (n=5), fructose experience (n=6), or glucose + fructose experience(n=5). (*:p<0.05, **:p<0.01,***:p<0.001,****:p<0.0001). All tests were conducted in the Davis Rig. Statistical Analysis are in Supplementary Table 3.

    Article Snippet: Briefly, mice were anesthetized with isoflurane (5% induction rate; 2-3% maintenance rate, as needed) to receive a scrambled sequence shRNA (Control, sc-108080, Santa Cruz Biotechnology), GCK shRNA (sc-35459-V, Santa Cruz Biotechnology), or Maltase-Glucoamylase shRNA (m) (sc-75741-V, Santa Cruz Biotechnology).

    Techniques: shRNA, Control, Concentration Assay, Expressing

    a Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Mean relative transcript expression of Mgam, Gck, and Tas1r3 in control (n= 4) and MGAM KD (n=4) in TRPM5+ mice c Schematic of burst structure (period of continuous licks separated by <1s) and time frame d Schematic of 300 lick test of 0.6M maltose in lickometer. e Schematical graph showing relationship between burst size and hedonic value. f Mean (±SEM) burst size, burst number, total licks for 300 lick test in control (n=3) and MGAM KD (n=3) in TRPM5+ mice (*:p<0.05). All tests were conducted in the gustometer. Statistical Analysis are in .

    Journal: bioRxiv

    Article Title: An enzymatic–metabolic sensing axis in taste cells detects glucose-yielding carbohydrates

    doi: 10.64898/2026.03.13.710876

    Figure Lengend Snippet: a Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Mean relative transcript expression of Mgam, Gck, and Tas1r3 in control (n= 4) and MGAM KD (n=4) in TRPM5+ mice c Schematic of burst structure (period of continuous licks separated by <1s) and time frame d Schematic of 300 lick test of 0.6M maltose in lickometer. e Schematical graph showing relationship between burst size and hedonic value. f Mean (±SEM) burst size, burst number, total licks for 300 lick test in control (n=3) and MGAM KD (n=3) in TRPM5+ mice (*:p<0.05). All tests were conducted in the gustometer. Statistical Analysis are in .

    Article Snippet: Briefly, mice were anesthetized with isoflurane (5% induction rate; 2-3% maintenance rate, as needed) to receive a scrambled sequence shRNA (Control, sc-108080, Santa Cruz Biotechnology), GCK shRNA (sc-35459-V, Santa Cruz Biotechnology), or Maltase-Glucoamylase shRNA (m) (sc-75741-V, Santa Cruz Biotechnology).

    Techniques: Control, shRNA, Expressing

    a Schematic depicting T1R2+T1R3 sweet receptor transgenic knockout mouse. b Mean transcript expression of Gck and Mgam for sugar naive (n=4) and sugar exposed (n=4) T1RKO. c Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. d Schematic of brief access taste test of maltose and sucrose in lickometer. e Block 1 mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=5) and MGAM KD (n=5) T1RKO. f Mean lick scores averaged across concentration for control and MGAM KD for first block (5/group). g Schematic of brief access taste test of glucose and fructose in lickometer. h First block mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M glucose and fructose in control (n=6) and MGAM KD (n=5) T1RKO. i First block glucose and fructose mean lick scores averaged across concentration for control (n=6) and MGAM KD (n=5) T1RKO. All tests were conducted in the Davis Rig. (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). Statistical Analysis are in Supplementary Table 7.

    Journal: bioRxiv

    Article Title: An enzymatic–metabolic sensing axis in taste cells detects glucose-yielding carbohydrates

    doi: 10.64898/2026.03.13.710876

    Figure Lengend Snippet: a Schematic depicting T1R2+T1R3 sweet receptor transgenic knockout mouse. b Mean transcript expression of Gck and Mgam for sugar naive (n=4) and sugar exposed (n=4) T1RKO. c Schematic of control and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. d Schematic of brief access taste test of maltose and sucrose in lickometer. e Block 1 mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M maltose and sucrose in control (n=5) and MGAM KD (n=5) T1RKO. f Mean lick scores averaged across concentration for control and MGAM KD for first block (5/group). g Schematic of brief access taste test of glucose and fructose in lickometer. h First block mean (±SEM) lick scores for 0.316M, 0.56M, 1.1M glucose and fructose in control (n=6) and MGAM KD (n=5) T1RKO. i First block glucose and fructose mean lick scores averaged across concentration for control (n=6) and MGAM KD (n=5) T1RKO. All tests were conducted in the Davis Rig. (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). Statistical Analysis are in Supplementary Table 7.

    Article Snippet: Briefly, mice were anesthetized with isoflurane (5% induction rate; 2-3% maintenance rate, as needed) to receive a scrambled sequence shRNA (Control, sc-108080, Santa Cruz Biotechnology), GCK shRNA (sc-35459-V, Santa Cruz Biotechnology), or Maltase-Glucoamylase shRNA (m) (sc-75741-V, Santa Cruz Biotechnology).

    Techniques: Transgenic Assay, Knock-Out, Expressing, Control, shRNA, Blocking Assay, Concentration Assay

    a Schematic showing control, Gck, and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Schematic of 1000 lick test of mixed nutrient solution (Peptamen) in lickometer for pre-surgery and post-surgery. c Mean total licks for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. d Mean meal duration for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. e Mean burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. f Mean burst number for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. g Mean (±SEM) cumulative first 15 burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. (n=12/group for all tests) (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). All tests were conducted in the gustometer. Statistical Analysis are in Supplementary Table 8.

    Journal: bioRxiv

    Article Title: An enzymatic–metabolic sensing axis in taste cells detects glucose-yielding carbohydrates

    doi: 10.64898/2026.03.13.710876

    Figure Lengend Snippet: a Schematic showing control, Gck, and Mgam virogenetic shRNA silencing surgery in the major taste fields of the tongue. b Schematic of 1000 lick test of mixed nutrient solution (Peptamen) in lickometer for pre-surgery and post-surgery. c Mean total licks for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. d Mean meal duration for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. e Mean burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. f Mean burst number for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. g Mean (±SEM) cumulative first 15 burst size for 1000 lick test in control, GCK KD, MGAM KD pre-test vs post-test. (n=12/group for all tests) (*:p<0.05, **:p<0.01, ***:p<0.001, ****:p<0.0001). All tests were conducted in the gustometer. Statistical Analysis are in Supplementary Table 8.

    Article Snippet: Briefly, mice were anesthetized with isoflurane (5% induction rate; 2-3% maintenance rate, as needed) to receive a scrambled sequence shRNA (Control, sc-108080, Santa Cruz Biotechnology), GCK shRNA (sc-35459-V, Santa Cruz Biotechnology), or Maltase-Glucoamylase shRNA (m) (sc-75741-V, Santa Cruz Biotechnology).

    Techniques: Control, shRNA

    L-plastin downregulation decreases the number of cells connected by TNTs. ( a ) Representative stain free gel is shown as a loading control. ( b ) Representative fluorescent Western blot demonstrating successful downregulation of L-plastin protein levels in shRNA-L-plastin cells compared to the shRNA-Control. ( c ) Quantification of L-plastin downregulation from Western blots shows a 77.2% downregulation in L-plastin expression (22.8 ± 6.48) compared to control cells. Data are the average of 5 independent experiments. Graph show means (±s.e.m; p = 0.0121). ( d ) Top Z-stack representative 3D image of control shRNA transduced cells and ( e ) Zoom in of ROI outlined in (dashed rectangles) ( d ). ( f ) Representative 3D image of shRNA L-plastin transduced cells and ( g ) Zoom in of ROI outlined in (dashed rectangles) ( f ). TNTs are highlighted with white arrowheads and cells connected by TNTs are identified with a white star (*). Scale bars for ( d , f ) = 10 μm and 5 μm for ( e , g ). Scale bars = 5 μm. ( h ) Down regulation of L-plastin resulted in a significantly lower percentage of cells connected by TNT (32.7 ± 4.7%) compared to the shRNA-control cells (51.2 ± 3.8%). The quantification of cells connected by TNTs came from 5 blinded independent experiments ( n = 520 for shRNA Ctl and n = 508 for shRNA L-plastin). Graph show means (±s.e.m; p = 0.0214). * p < 0.05.

    Journal: Cells

    Article Title: Myosin-X Acts Upstream of L-Plastin to Drive Stress-Induced Tunneling Nanotubes

    doi: 10.3390/cells15030224

    Figure Lengend Snippet: L-plastin downregulation decreases the number of cells connected by TNTs. ( a ) Representative stain free gel is shown as a loading control. ( b ) Representative fluorescent Western blot demonstrating successful downregulation of L-plastin protein levels in shRNA-L-plastin cells compared to the shRNA-Control. ( c ) Quantification of L-plastin downregulation from Western blots shows a 77.2% downregulation in L-plastin expression (22.8 ± 6.48) compared to control cells. Data are the average of 5 independent experiments. Graph show means (±s.e.m; p = 0.0121). ( d ) Top Z-stack representative 3D image of control shRNA transduced cells and ( e ) Zoom in of ROI outlined in (dashed rectangles) ( d ). ( f ) Representative 3D image of shRNA L-plastin transduced cells and ( g ) Zoom in of ROI outlined in (dashed rectangles) ( f ). TNTs are highlighted with white arrowheads and cells connected by TNTs are identified with a white star (*). Scale bars for ( d , f ) = 10 μm and 5 μm for ( e , g ). Scale bars = 5 μm. ( h ) Down regulation of L-plastin resulted in a significantly lower percentage of cells connected by TNT (32.7 ± 4.7%) compared to the shRNA-control cells (51.2 ± 3.8%). The quantification of cells connected by TNTs came from 5 blinded independent experiments ( n = 520 for shRNA Ctl and n = 508 for shRNA L-plastin). Graph show means (±s.e.m; p = 0.0214). * p < 0.05.

    Article Snippet: For down regulation experiments, Myo10 shRNA (SCBT, sc-43242-V), L-plastin shRNA (SCBT, sc-43209-V), control shRNA lentiviral particles (SCBT, Dallas, TX, USA, sc-108060), Polybrene (sc-134220) reagent and puromycin dihydrochloride (Santa Cruz, sc-108071) were from Santa Cruz Biotechnology, Inc., and were used according to the manufacturer’s instructions.

    Techniques: Staining, Control, Western Blot, shRNA, Expressing

    L-plastin expression levels dos not affect Myo10 expression. ( a ) Representative stain free gel is shown as a loading control. ( b ) Representative fluorescent Western blot showing up-regulation of L-plastin expression and its effect on Myo10 protein levels. ( c ) Quantitative analysis showing no statistical significant changes (ns) in Myo10 expression (92.8% ± 2.3%; p = 0.9203) upon L-plastin upregulation. ( d , e ) Stain free gel and Western blot from a,b showing successful down-regulation of L-plastin (~70 KDa) in ShRNA L-plastin cells. To see its effect on Myo10 expression, the top part of the blot was probed with Myo10 antibody to determine the effect of L-plastin down regulation on Myo10 protein levels (~250 KDa). ( f ) Quantification of relative levels of Myo 10 expression shows no significant changes (96.4%; (ns) with p = 0.1443) compared to control cells. Data are the average of 5 independent experiments, and the graph shows means (±s.e.m).

    Journal: Cells

    Article Title: Myosin-X Acts Upstream of L-Plastin to Drive Stress-Induced Tunneling Nanotubes

    doi: 10.3390/cells15030224

    Figure Lengend Snippet: L-plastin expression levels dos not affect Myo10 expression. ( a ) Representative stain free gel is shown as a loading control. ( b ) Representative fluorescent Western blot showing up-regulation of L-plastin expression and its effect on Myo10 protein levels. ( c ) Quantitative analysis showing no statistical significant changes (ns) in Myo10 expression (92.8% ± 2.3%; p = 0.9203) upon L-plastin upregulation. ( d , e ) Stain free gel and Western blot from a,b showing successful down-regulation of L-plastin (~70 KDa) in ShRNA L-plastin cells. To see its effect on Myo10 expression, the top part of the blot was probed with Myo10 antibody to determine the effect of L-plastin down regulation on Myo10 protein levels (~250 KDa). ( f ) Quantification of relative levels of Myo 10 expression shows no significant changes (96.4%; (ns) with p = 0.1443) compared to control cells. Data are the average of 5 independent experiments, and the graph shows means (±s.e.m).

    Article Snippet: For down regulation experiments, Myo10 shRNA (SCBT, sc-43242-V), L-plastin shRNA (SCBT, sc-43209-V), control shRNA lentiviral particles (SCBT, Dallas, TX, USA, sc-108060), Polybrene (sc-134220) reagent and puromycin dihydrochloride (Santa Cruz, sc-108071) were from Santa Cruz Biotechnology, Inc., and were used according to the manufacturer’s instructions.

    Techniques: Expressing, Staining, Control, Western Blot, shRNA

    Myo10 expression levels affect L-plastin expression. ( a ) Representative stain free gel is shown as a loading control. ( b ) Representative fluorescent Western blot showing that down-regulation of Myo10 expression (61.0% ± 2.4% with p = 0.0121) using shRNA lentiviral particles ( c ) results in the reduction of L-plastin expression levels to 68% (±2.8%; p -value: 0.0121). Data are the average of 5 independent experiments. ( d ) Conversely, we looked at the effects of up-regulation of Myo10 using transient GFP-Myo10 transfection. ( e ) Representative stain free gel is shown as a loading control. To avoid that the very bright GFP signal affects the endogenous Myo10 signal, the GFP-vector experiments were loaded first, followed by the GFP-Myo10 transfected cells on the same gels. Here a representative experiment is shown, where the control lane (GFP-vector) and its matching experimental lane (GFP-Myo10) from the same gel were juxtaposed as shown by the black separation lane. The entire gel is shown in ; lanes 1 ( e ) and 4 ( f ) were used as a representative image here. ( f ) Representative fluorescent Western blot of the same samples as ( e ) are shown and it demonstrates that up-regulation of GFP-Myo10 expression results in the upregulation of endogenous L-plastin expression. ( g ) Quantitative analysis shows a 132.3.3% (±9.4%; p = 0.0121) increase in L-plastin in cells over-expressing GFP-Myo10 compared to the control GFP expressing cells. Data are the average of 5 independent experiments. * p < 0.05.

    Journal: Cells

    Article Title: Myosin-X Acts Upstream of L-Plastin to Drive Stress-Induced Tunneling Nanotubes

    doi: 10.3390/cells15030224

    Figure Lengend Snippet: Myo10 expression levels affect L-plastin expression. ( a ) Representative stain free gel is shown as a loading control. ( b ) Representative fluorescent Western blot showing that down-regulation of Myo10 expression (61.0% ± 2.4% with p = 0.0121) using shRNA lentiviral particles ( c ) results in the reduction of L-plastin expression levels to 68% (±2.8%; p -value: 0.0121). Data are the average of 5 independent experiments. ( d ) Conversely, we looked at the effects of up-regulation of Myo10 using transient GFP-Myo10 transfection. ( e ) Representative stain free gel is shown as a loading control. To avoid that the very bright GFP signal affects the endogenous Myo10 signal, the GFP-vector experiments were loaded first, followed by the GFP-Myo10 transfected cells on the same gels. Here a representative experiment is shown, where the control lane (GFP-vector) and its matching experimental lane (GFP-Myo10) from the same gel were juxtaposed as shown by the black separation lane. The entire gel is shown in ; lanes 1 ( e ) and 4 ( f ) were used as a representative image here. ( f ) Representative fluorescent Western blot of the same samples as ( e ) are shown and it demonstrates that up-regulation of GFP-Myo10 expression results in the upregulation of endogenous L-plastin expression. ( g ) Quantitative analysis shows a 132.3.3% (±9.4%; p = 0.0121) increase in L-plastin in cells over-expressing GFP-Myo10 compared to the control GFP expressing cells. Data are the average of 5 independent experiments. * p < 0.05.

    Article Snippet: For down regulation experiments, Myo10 shRNA (SCBT, sc-43242-V), L-plastin shRNA (SCBT, sc-43209-V), control shRNA lentiviral particles (SCBT, Dallas, TX, USA, sc-108060), Polybrene (sc-134220) reagent and puromycin dihydrochloride (Santa Cruz, sc-108071) were from Santa Cruz Biotechnology, Inc., and were used according to the manufacturer’s instructions.

    Techniques: Expressing, Staining, Control, Western Blot, shRNA, Transfection, Plasmid Preparation