rapa Search Results


rapamycin  (Alomone Labs)
90
Alomone Labs rapamycin
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Rapamycin, supplied by Alomone Labs, 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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standard sirolimus d3  (Toronto Research Chemicals)
91
Toronto Research Chemicals standard sirolimus d3
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Standard Sirolimus D3, supplied by Toronto Research Chemicals, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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brassica rapa  (Carolina Biological)
92
Carolina Biological brassica rapa
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Brassica Rapa, supplied by Carolina Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fast cycling brassica rapa  (Carolina Biological)
91
Carolina Biological fast cycling brassica rapa
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Fast Cycling Brassica Rapa, supplied by Carolina Biological, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rapeseed oil  (Valiant Co Ltd)
96
Valiant Co Ltd rapeseed oil
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Rapeseed Oil, supplied by Valiant Co Ltd, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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autophagy inhibitor 3 ma group  (Proteintech)
92
Proteintech autophagy inhibitor 3 ma group
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Autophagy Inhibitor 3 Ma Group, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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brassica rapa complete rdna  (Biotechnology Information)
90
Biotechnology Information brassica rapa complete rdna
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Brassica Rapa Complete Rdna, supplied by Biotechnology Information, 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
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seeds of the sulforaphane-rich broccoletti prototype brassica rapa sylvestris  (Megerle Online)
90
Megerle Online seeds of the sulforaphane-rich broccoletti prototype brassica rapa sylvestris
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Seeds Of The Sulforaphane Rich Broccoletti Prototype Brassica Rapa Sylvestris, supplied by Megerle Online, 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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rapa chemical  (Sinopharm ltd)
90
Sinopharm ltd rapa chemical
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Rapa Chemical, supplied by Sinopharm ltd, 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
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purple pak-choi  (Makino Inc)
90
Makino Inc purple pak-choi
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Purple Pak Choi, supplied by Makino Inc, 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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brassica rapa  (Siemens AG)
90
Siemens AG brassica rapa
Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. <t>Rapamycin</t> (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.
Brassica Rapa, supplied by Siemens AG, 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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Image Search Results


Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. Rapamycin (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Dual inhibition of BDNF/TrkB and autophagy: a promising therapeutic approach for colorectal cancer

doi: 10.1111/jcmm.13181

Figure Lengend Snippet: Effect of tyrosine kinase receptor inhibitor (K252a) on TrkB/BDNF expression and autophagy induction. SW480 and SW620 were treated with 100 nM of K252a for 3 hrs, recovered, washed and total RNA and proteins were extracted (see ). ( A ) BDNF, Full Length (FL) and Truncated (Tr) TrkB, transcripts expression was assessed by qPCR. Histograms are representative of three independent experiments. The fold expression was obtained by the comparative cycle threshold method using the GAPDH RNA expression level as internal control. ( B ) BDNF, FL TrkB, Tr TrkB, phospho‐TrkB (P TrkB), AKT, phospho‐AKT (P AKT), mTOR and phospho‐mTOR (P mTOR), proteins expression were assessed by Western blotting. The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( C ) Staining of SW480 and SW620 was realized with anti‐BDNF antibody (green), anti‐TrkB antibody (red) and DAPI (blue). Images were obtained through confocal microscopy (magnification ×1000). ( D ) Beclin1, ATG5 and LC3 proteins expression was assessed by Western blotting after treatment with either K252a (100 nM, 3 hrs) or with CQ (25 μM, 3 hrs). The density of each band representative for protein expression was calculated with ImageJ software. Images show representative results of three experiments. Statistically results are explained in comparison with control cells, normalized at 1 (* P < 0.05; ** P < 0.01; *** P < 0.001). ( E ) Staining of SW480 and SW620 was realized with anti‐LC3 antibody (green) and DAPI (blue) through indirect immunofluorescence. Rapamycin (20 nM, 3 hrs) was used as a positive control for autophagy induction. Relative fluorescence quantification was accomplished by using a green surface plot with the ImageJ software application.

Article Snippet: K252a (Alomone Labs, Jerusalem, Israel), chloroquine (CQ) and rapamycin (Sigma‐Aldrich, St. Quentin Fallavier, France) were used for NT, autophagy and mTOR inhibition, respectively.

Techniques: Expressing, RNA Expression, Western Blot, Software, Staining, Confocal Microscopy, Immunofluorescence, Positive Control, Fluorescence