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lambda protein phosphatase 100 u/ml  (Beyotime)
90
Beyotime lambda protein phosphatase 100 u/ml
a , b HK-2 cells transfected with siNEU1 ( a ) or NEU1 full-length plasmid ( b ) for 24 h and treated with TGFβ (10 ng/ml) for 24 h. Then the cells were incubated with cycloheximide (CHX, 20 μg/ml) for the indicated periods of time (0, 2, 4, 8, 12, 24 h) (left). Lysates were harvested from the cells and analyzed by western blots. Quantitation of ALK5 protein levels were shown in the right pane. n = 3 samples per group. Unpaired t -test. c – f Western blots ( c , e ) and quantitative results ( d , f ) of p-ALK5, ALK5, p-SMAD2/3, and SMAD2/3 in HK-2 cells transfected with siNEU1 or NEU1 plasmid for 24 h and treated with TGFβ (10 ng/ml) for 24 h. n = 3 samples per group. Relative protein levels were shown after normalization to GAPDH. One-way ANOVA followed by Tukey’s multiple comparisons test. g , h Western blots ( g ) and quantitative results ( h ) of p-ALK5, p-SMAD2/3, and SMAD2/3 in HK-2 cells transfected with NEU1 and ALK5 160-200 plasmid for 24 h and treated with TGFβ (10 ng/ml) for 24 h. n = 3 samples per group. Relative protein levels were shown after normalization to GAPDH. One-way ANOVA followed by Tukey’s multiple comparisons test. i Co-immunoprecipitation of NEU1 and ALK5 in HK-2 cells stimulated with TGFβ (10 ng/ml) for 24 h. <t>λpp,</t> <t>Lambda</t> Protein <t>Phosphatase.</t> Two independent experiments were performed. j Western blots of ALK5 in HK-2 cells. siALK5-1, siALK5-2, and siALK5-3 were 3 different siRNA sequences. HK-2 cells were transduced with siALK5 for 48 h. Two independent experiments were performed. k – m mRNA levels of KIM1 ( k ) and protein level of p-ALK5, p-SMAD2/3, and SMAD2/3 ( l , m ) in HK-2 cells transduced with NEU1 full-length plasmid and ALK5 siRNA for 48 h. KIM1 mRNA normalized to GAPDH . Relative protein levels were shown after normalization to GAPDH. k n = 3 samples per group. l , m n = 3 samples per group. Two-way ANOVA followed by Tukey’s multiple comparisons test. n The proposed mechanisms of NEU1-mediated renal fibrosis. All statistic data were presented as mean ± SD. All tests were two-tailed.
Lambda Protein Phosphatase 100 U/Ml, supplied by Beyotime, 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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Average 90 stars, based on 1 article reviews
lambda protein phosphatase 100 u/ml - by Bioz Stars, 2026-03
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a , b HK-2 cells transfected with siNEU1 ( a ) or NEU1 full-length plasmid ( b ) for 24 h and treated with TGFβ (10 ng/ml) for 24 h. Then the cells were incubated with cycloheximide (CHX, 20 μg/ml) for the indicated periods of time (0, 2, 4, 8, 12, 24 h) (left). Lysates were harvested from the cells and analyzed by western blots. Quantitation of ALK5 protein levels were shown in the right pane. n = 3 samples per group. Unpaired t -test. c – f Western blots ( c , e ) and quantitative results ( d , f ) of p-ALK5, ALK5, p-SMAD2/3, and SMAD2/3 in HK-2 cells transfected with siNEU1 or NEU1 plasmid for 24 h and treated with TGFβ (10 ng/ml) for 24 h. n = 3 samples per group. Relative protein levels were shown after normalization to GAPDH. One-way ANOVA followed by Tukey’s multiple comparisons test. g , h Western blots ( g ) and quantitative results ( h ) of p-ALK5, p-SMAD2/3, and SMAD2/3 in HK-2 cells transfected with NEU1 and ALK5 160-200 plasmid for 24 h and treated with TGFβ (10 ng/ml) for 24 h. n = 3 samples per group. Relative protein levels were shown after normalization to GAPDH. One-way ANOVA followed by Tukey’s multiple comparisons test. i Co-immunoprecipitation of NEU1 and ALK5 in HK-2 cells stimulated with TGFβ (10 ng/ml) for 24 h. λpp, Lambda Protein Phosphatase. Two independent experiments were performed. j Western blots of ALK5 in HK-2 cells. siALK5-1, siALK5-2, and siALK5-3 were 3 different siRNA sequences. HK-2 cells were transduced with siALK5 for 48 h. Two independent experiments were performed. k – m mRNA levels of KIM1 ( k ) and protein level of p-ALK5, p-SMAD2/3, and SMAD2/3 ( l , m ) in HK-2 cells transduced with NEU1 full-length plasmid and ALK5 siRNA for 48 h. KIM1 mRNA normalized to GAPDH . Relative protein levels were shown after normalization to GAPDH. k n = 3 samples per group. l , m n = 3 samples per group. Two-way ANOVA followed by Tukey’s multiple comparisons test. n The proposed mechanisms of NEU1-mediated renal fibrosis. All statistic data were presented as mean ± SD. All tests were two-tailed.

Journal: Nature Communications

Article Title: Neuraminidase 1 promotes renal fibrosis development in male mice

doi: 10.1038/s41467-023-37450-8

Figure Lengend Snippet: a , b HK-2 cells transfected with siNEU1 ( a ) or NEU1 full-length plasmid ( b ) for 24 h and treated with TGFβ (10 ng/ml) for 24 h. Then the cells were incubated with cycloheximide (CHX, 20 μg/ml) for the indicated periods of time (0, 2, 4, 8, 12, 24 h) (left). Lysates were harvested from the cells and analyzed by western blots. Quantitation of ALK5 protein levels were shown in the right pane. n = 3 samples per group. Unpaired t -test. c – f Western blots ( c , e ) and quantitative results ( d , f ) of p-ALK5, ALK5, p-SMAD2/3, and SMAD2/3 in HK-2 cells transfected with siNEU1 or NEU1 plasmid for 24 h and treated with TGFβ (10 ng/ml) for 24 h. n = 3 samples per group. Relative protein levels were shown after normalization to GAPDH. One-way ANOVA followed by Tukey’s multiple comparisons test. g , h Western blots ( g ) and quantitative results ( h ) of p-ALK5, p-SMAD2/3, and SMAD2/3 in HK-2 cells transfected with NEU1 and ALK5 160-200 plasmid for 24 h and treated with TGFβ (10 ng/ml) for 24 h. n = 3 samples per group. Relative protein levels were shown after normalization to GAPDH. One-way ANOVA followed by Tukey’s multiple comparisons test. i Co-immunoprecipitation of NEU1 and ALK5 in HK-2 cells stimulated with TGFβ (10 ng/ml) for 24 h. λpp, Lambda Protein Phosphatase. Two independent experiments were performed. j Western blots of ALK5 in HK-2 cells. siALK5-1, siALK5-2, and siALK5-3 were 3 different siRNA sequences. HK-2 cells were transduced with siALK5 for 48 h. Two independent experiments were performed. k – m mRNA levels of KIM1 ( k ) and protein level of p-ALK5, p-SMAD2/3, and SMAD2/3 ( l , m ) in HK-2 cells transduced with NEU1 full-length plasmid and ALK5 siRNA for 48 h. KIM1 mRNA normalized to GAPDH . Relative protein levels were shown after normalization to GAPDH. k n = 3 samples per group. l , m n = 3 samples per group. Two-way ANOVA followed by Tukey’s multiple comparisons test. n The proposed mechanisms of NEU1-mediated renal fibrosis. All statistic data were presented as mean ± SD. All tests were two-tailed.

Article Snippet: The cell lysis solution was incubated with lambda protein phosphatase (λpp, 100 U/ml, Beyotime) and MnCl 2 (1 mM) for 30 min at 30 °C.

Techniques: Transfection, Plasmid Preparation, Incubation, Western Blot, Quantitation Assay, Immunoprecipitation, Transduction, Two Tailed Test