proteinase k Thermo Fisher Search Results


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  • 99
    New England Biolabs proteinase k
    Proteinase K, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 4094 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher proteinase k solution
    Spectrophotometry analysis of pig LV biopsy RNA. RNA extracted according to each manufacturer’s protocol, with the addition of a <t>proteinase</t> K digestion in half the samples. n = 3 biopsies/experimental condition, mean±SEM. N . B . Exiqon miRCURY tissue protocol requires proteinase K, therefore no–proteinase K condition was done with this kit. (A) RNA yield, assessed as ng RNA recovered per mg input tissue, was not significantly different between the protocols (Kruskal-Wallis test with Dunn correction), and additional proteinase K digestion in the RNAqueous micro, mir Vana and miRNeasy micro protocols did not significantly improve RNA yield (Kruskal-Wallis test with Dunn correction). (B) 260/280 and 260/230 ratios provide an assessment of the purity of the RNA. Ratio
    Proteinase K Solution, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1552 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher rna grade proteinase k
    ER stress is activated in prion disease. Upregulation of ER stress markers was determined in prion infected CD1 mice (n = 2 per timepoint) after inoculation with 6.5logLD 50 RML prions. Western blot analysis was performed to analyze the levels of C12, Grp58, JNK phosphorylation, ERK phosphorylation, total PrP and proteinase K-resistant PrP. The total level of JNK, ERK, and actin were measured as loading controls. Faint processing of C12 is visible at 4 months post inoculation (mpi) but more clearly at 4.5 and 5 mpi (active fragments of C12 are indicated by an arrow head). Phosphorylation of JNK (P-JNK) as well as induction of Grp58 was observed at 4.5 and 5 mpi. Phosphorylation of ERK was observed at 4.5 and 5 mpi. Higher order SDS-resistant PrP species were first visible at 3 mpi and thereafter (an arrow head marks the migration of monomeric PrP) along with <t>proteinase</t> K resistant PrP.
    Rna Grade Proteinase K, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2640 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna grade proteinase k/product/Thermo Fisher
    Average 99 stars, based on 2640 article reviews
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    rna grade proteinase k - by Bioz Stars, 2020-07
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    99
    Qiagen proteinase k
    ER stress is activated in prion disease. Upregulation of ER stress markers was determined in prion infected CD1 mice (n = 2 per timepoint) after inoculation with 6.5logLD 50 RML prions. Western blot analysis was performed to analyze the levels of C12, Grp58, JNK phosphorylation, ERK phosphorylation, total PrP and proteinase K-resistant PrP. The total level of JNK, ERK, and actin were measured as loading controls. Faint processing of C12 is visible at 4 months post inoculation (mpi) but more clearly at 4.5 and 5 mpi (active fragments of C12 are indicated by an arrow head). Phosphorylation of JNK (P-JNK) as well as induction of Grp58 was observed at 4.5 and 5 mpi. Phosphorylation of ERK was observed at 4.5 and 5 mpi. Higher order SDS-resistant PrP species were first visible at 3 mpi and thereafter (an arrow head marks the migration of monomeric PrP) along with <t>proteinase</t> K resistant PrP.
    Proteinase K, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 20858 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher proteinase k powder
    GORASP2 is localized to autophagosomes and lysosomes upon starvation. ( a ) GORASP2 colocalizes with GFP-LC3 upon starvation. GFP-LC3 HeLa cells were treated with growth medium (ctr), BafA1, EBSS or EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, GOLGA2 and DNA. The bottom row shows higher magnifications of the indicated area in the above row. Scale bar: 10 µm in the upper four rows, 3 µm in the bottom row. ( b ) Quantification of ( a ) for the percentage of GORASP2 puncta that colocalized with GFP-LC3. ( c ) GORASP2 colocalizes with LC3 and LAMP2 but not EEA1 or LAMP1 upon starvation. HeLa cells were treated with EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, LC3, EEA1, LAMP1, or LAMP2 and DNA as indicated. The three rows on the right are higher magnifications of the boxed area in the three rows on the left. Scale bars: 10 µm in the left rows, 3 µm in the right rows. ( d ) Quantification of ( c ) for the percentage of GORASP2 puncta that colocalized with LC3, EEA1, LAMP1 or LAMP2. ( e ) Western blot of the <t>proteinase</t> K protection assay. HeLa cells were treated with growth medium (ctr) or EBSS and 400 nM BafA1 (E + B) for 4 h, then the collected PNS were equally divided into three tubes, one left untreated, one was incubated with 2.5 μg/ml protease K (PK) only, and one was treated with both PK and 1% TritonX-100 (TX-100) for 10 min.
    Proteinase K Powder, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Thermo Fisher en0531 protease k
    GORASP2 is localized to autophagosomes and lysosomes upon starvation. ( a ) GORASP2 colocalizes with GFP-LC3 upon starvation. GFP-LC3 HeLa cells were treated with growth medium (ctr), BafA1, EBSS or EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, GOLGA2 and DNA. The bottom row shows higher magnifications of the indicated area in the above row. Scale bar: 10 µm in the upper four rows, 3 µm in the bottom row. ( b ) Quantification of ( a ) for the percentage of GORASP2 puncta that colocalized with GFP-LC3. ( c ) GORASP2 colocalizes with LC3 and LAMP2 but not EEA1 or LAMP1 upon starvation. HeLa cells were treated with EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, LC3, EEA1, LAMP1, or LAMP2 and DNA as indicated. The three rows on the right are higher magnifications of the boxed area in the three rows on the left. Scale bars: 10 µm in the left rows, 3 µm in the right rows. ( d ) Quantification of ( c ) for the percentage of GORASP2 puncta that colocalized with LC3, EEA1, LAMP1 or LAMP2. ( e ) Western blot of the <t>proteinase</t> K protection assay. HeLa cells were treated with growth medium (ctr) or EBSS and 400 nM BafA1 (E + B) for 4 h, then the collected PNS were equally divided into three tubes, one left untreated, one was incubated with 2.5 μg/ml protease K (PK) only, and one was treated with both PK and 1% TritonX-100 (TX-100) for 10 min.
    En0531 Protease K, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/en0531 protease k/product/Thermo Fisher
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    97
    Thermo Fisher proteinase k solution pbs
    GORASP2 is localized to autophagosomes and lysosomes upon starvation. ( a ) GORASP2 colocalizes with GFP-LC3 upon starvation. GFP-LC3 HeLa cells were treated with growth medium (ctr), BafA1, EBSS or EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, GOLGA2 and DNA. The bottom row shows higher magnifications of the indicated area in the above row. Scale bar: 10 µm in the upper four rows, 3 µm in the bottom row. ( b ) Quantification of ( a ) for the percentage of GORASP2 puncta that colocalized with GFP-LC3. ( c ) GORASP2 colocalizes with LC3 and LAMP2 but not EEA1 or LAMP1 upon starvation. HeLa cells were treated with EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, LC3, EEA1, LAMP1, or LAMP2 and DNA as indicated. The three rows on the right are higher magnifications of the boxed area in the three rows on the left. Scale bars: 10 µm in the left rows, 3 µm in the right rows. ( d ) Quantification of ( c ) for the percentage of GORASP2 puncta that colocalized with LC3, EEA1, LAMP1 or LAMP2. ( e ) Western blot of the <t>proteinase</t> K protection assay. HeLa cells were treated with growth medium (ctr) or EBSS and 400 nM BafA1 (E + B) for 4 h, then the collected PNS were equally divided into three tubes, one left untreated, one was incubated with 2.5 μg/ml protease K (PK) only, and one was treated with both PK and 1% TritonX-100 (TX-100) for 10 min.
    Proteinase K Solution Pbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/proteinase k solution pbs/product/Thermo Fisher
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    99
    Millipore proteinase k
    GORASP2 is localized to autophagosomes and lysosomes upon starvation. ( a ) GORASP2 colocalizes with GFP-LC3 upon starvation. GFP-LC3 HeLa cells were treated with growth medium (ctr), BafA1, EBSS or EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, GOLGA2 and DNA. The bottom row shows higher magnifications of the indicated area in the above row. Scale bar: 10 µm in the upper four rows, 3 µm in the bottom row. ( b ) Quantification of ( a ) for the percentage of GORASP2 puncta that colocalized with GFP-LC3. ( c ) GORASP2 colocalizes with LC3 and LAMP2 but not EEA1 or LAMP1 upon starvation. HeLa cells were treated with EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, LC3, EEA1, LAMP1, or LAMP2 and DNA as indicated. The three rows on the right are higher magnifications of the boxed area in the three rows on the left. Scale bars: 10 µm in the left rows, 3 µm in the right rows. ( d ) Quantification of ( c ) for the percentage of GORASP2 puncta that colocalized with LC3, EEA1, LAMP1 or LAMP2. ( e ) Western blot of the <t>proteinase</t> K protection assay. HeLa cells were treated with growth medium (ctr) or EBSS and 400 nM BafA1 (E + B) for 4 h, then the collected PNS were equally divided into three tubes, one left untreated, one was incubated with 2.5 μg/ml protease K (PK) only, and one was treated with both PK and 1% TritonX-100 (TX-100) for 10 min.
    Proteinase K, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 26156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher pcr grade proteinase k solution
    GORASP2 is localized to autophagosomes and lysosomes upon starvation. ( a ) GORASP2 colocalizes with GFP-LC3 upon starvation. GFP-LC3 HeLa cells were treated with growth medium (ctr), BafA1, EBSS or EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, GOLGA2 and DNA. The bottom row shows higher magnifications of the indicated area in the above row. Scale bar: 10 µm in the upper four rows, 3 µm in the bottom row. ( b ) Quantification of ( a ) for the percentage of GORASP2 puncta that colocalized with GFP-LC3. ( c ) GORASP2 colocalizes with LC3 and LAMP2 but not EEA1 or LAMP1 upon starvation. HeLa cells were treated with EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, LC3, EEA1, LAMP1, or LAMP2 and DNA as indicated. The three rows on the right are higher magnifications of the boxed area in the three rows on the left. Scale bars: 10 µm in the left rows, 3 µm in the right rows. ( d ) Quantification of ( c ) for the percentage of GORASP2 puncta that colocalized with LC3, EEA1, LAMP1 or LAMP2. ( e ) Western blot of the <t>proteinase</t> K protection assay. HeLa cells were treated with growth medium (ctr) or EBSS and 400 nM BafA1 (E + B) for 4 h, then the collected PNS were equally divided into three tubes, one left untreated, one was incubated with 2.5 μg/ml protease K (PK) only, and one was treated with both PK and 1% TritonX-100 (TX-100) for 10 min.
    Pcr Grade Proteinase K Solution, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Spectrophotometry analysis of pig LV biopsy RNA. RNA extracted according to each manufacturer’s protocol, with the addition of a proteinase K digestion in half the samples. n = 3 biopsies/experimental condition, mean±SEM. N . B . Exiqon miRCURY tissue protocol requires proteinase K, therefore no–proteinase K condition was done with this kit. (A) RNA yield, assessed as ng RNA recovered per mg input tissue, was not significantly different between the protocols (Kruskal-Wallis test with Dunn correction), and additional proteinase K digestion in the RNAqueous micro, mir Vana and miRNeasy micro protocols did not significantly improve RNA yield (Kruskal-Wallis test with Dunn correction). (B) 260/280 and 260/230 ratios provide an assessment of the purity of the RNA. Ratio

    Journal: PLoS ONE

    Article Title: Optimisation of laboratory methods for whole transcriptomic RNA analyses in human left ventricular biopsies and blood samples of clinical relevance

    doi: 10.1371/journal.pone.0213685

    Figure Lengend Snippet: Spectrophotometry analysis of pig LV biopsy RNA. RNA extracted according to each manufacturer’s protocol, with the addition of a proteinase K digestion in half the samples. n = 3 biopsies/experimental condition, mean±SEM. N . B . Exiqon miRCURY tissue protocol requires proteinase K, therefore no–proteinase K condition was done with this kit. (A) RNA yield, assessed as ng RNA recovered per mg input tissue, was not significantly different between the protocols (Kruskal-Wallis test with Dunn correction), and additional proteinase K digestion in the RNAqueous micro, mir Vana and miRNeasy micro protocols did not significantly improve RNA yield (Kruskal-Wallis test with Dunn correction). (B) 260/280 and 260/230 ratios provide an assessment of the purity of the RNA. Ratio

    Article Snippet: Thereafter, the manufacturer’s protocol was followed exactly in one set of extractions, while in a parallel set of extractions the protocol was modified to include a proteinase K digestion step; samples were incubated with 200 μg/mL proteinase K (Invitrogen AM2546) for 15 minutes at 55°C.

    Techniques: Spectrophotometry

    Summary of experimental design for Figs 2 , 3 and 4 . (A) Left ventricular biopsies were taken from three pigs undergoing cardio-pulmonary bypass (CPB). 7 biopsies were taken per pig. RNA was isolated from the biopsies using four commercially available kits; RNAqueous micro, mir Vana, miRCURY tissue, miRNeasy micro. Three kits (RNAqueous micro, mir Vana, miRNeasy micro) were tested with and without an additional proteinase K digestion step, giving 7 protocols in total. The miRCURY tissue protocol includes a proteinase K digestion, therefore this kit was not tested without this. RNA samples were characterised by spectrophotometry and capillary electrophoresis. (B) Blood samples were taken from three pigs undergoing CPB and processed to plasma. RNA was isolated from the plasma using four commercially-available kits; miRNeasy serum/plasma, mir Vana, miRCURY biofluids, Norgen Biotek plasma/serum. Each kit was tested with and without glycogen as a co-precipitant, giving 8 protocols in total. RNA samples were evaluated by RT-qPCR for the exogenous spike-in cel-miR-39-3p, and endogenous miRNAs miR-16-5p, miR-21-5p, miR-92a-3p.

    Journal: PLoS ONE

    Article Title: Optimisation of laboratory methods for whole transcriptomic RNA analyses in human left ventricular biopsies and blood samples of clinical relevance

    doi: 10.1371/journal.pone.0213685

    Figure Lengend Snippet: Summary of experimental design for Figs 2 , 3 and 4 . (A) Left ventricular biopsies were taken from three pigs undergoing cardio-pulmonary bypass (CPB). 7 biopsies were taken per pig. RNA was isolated from the biopsies using four commercially available kits; RNAqueous micro, mir Vana, miRCURY tissue, miRNeasy micro. Three kits (RNAqueous micro, mir Vana, miRNeasy micro) were tested with and without an additional proteinase K digestion step, giving 7 protocols in total. The miRCURY tissue protocol includes a proteinase K digestion, therefore this kit was not tested without this. RNA samples were characterised by spectrophotometry and capillary electrophoresis. (B) Blood samples were taken from three pigs undergoing CPB and processed to plasma. RNA was isolated from the plasma using four commercially-available kits; miRNeasy serum/plasma, mir Vana, miRCURY biofluids, Norgen Biotek plasma/serum. Each kit was tested with and without glycogen as a co-precipitant, giving 8 protocols in total. RNA samples were evaluated by RT-qPCR for the exogenous spike-in cel-miR-39-3p, and endogenous miRNAs miR-16-5p, miR-21-5p, miR-92a-3p.

    Article Snippet: Thereafter, the manufacturer’s protocol was followed exactly in one set of extractions, while in a parallel set of extractions the protocol was modified to include a proteinase K digestion step; samples were incubated with 200 μg/mL proteinase K (Invitrogen AM2546) for 15 minutes at 55°C.

    Techniques: Isolation, Spectrophotometry, Electrophoresis, Quantitative RT-PCR

    ER stress is activated in prion disease. Upregulation of ER stress markers was determined in prion infected CD1 mice (n = 2 per timepoint) after inoculation with 6.5logLD 50 RML prions. Western blot analysis was performed to analyze the levels of C12, Grp58, JNK phosphorylation, ERK phosphorylation, total PrP and proteinase K-resistant PrP. The total level of JNK, ERK, and actin were measured as loading controls. Faint processing of C12 is visible at 4 months post inoculation (mpi) but more clearly at 4.5 and 5 mpi (active fragments of C12 are indicated by an arrow head). Phosphorylation of JNK (P-JNK) as well as induction of Grp58 was observed at 4.5 and 5 mpi. Phosphorylation of ERK was observed at 4.5 and 5 mpi. Higher order SDS-resistant PrP species were first visible at 3 mpi and thereafter (an arrow head marks the migration of monomeric PrP) along with proteinase K resistant PrP.

    Journal: Prion

    Article Title: Prion Pathogenesis is Independent of Caspase-12

    doi:

    Figure Lengend Snippet: ER stress is activated in prion disease. Upregulation of ER stress markers was determined in prion infected CD1 mice (n = 2 per timepoint) after inoculation with 6.5logLD 50 RML prions. Western blot analysis was performed to analyze the levels of C12, Grp58, JNK phosphorylation, ERK phosphorylation, total PrP and proteinase K-resistant PrP. The total level of JNK, ERK, and actin were measured as loading controls. Faint processing of C12 is visible at 4 months post inoculation (mpi) but more clearly at 4.5 and 5 mpi (active fragments of C12 are indicated by an arrow head). Phosphorylation of JNK (P-JNK) as well as induction of Grp58 was observed at 4.5 and 5 mpi. Phosphorylation of ERK was observed at 4.5 and 5 mpi. Higher order SDS-resistant PrP species were first visible at 3 mpi and thereafter (an arrow head marks the migration of monomeric PrP) along with proteinase K resistant PrP.

    Article Snippet: For proteinase K treatment, 10% brain homogenates from terminally ill WT or Caspase-12 KOs were diluted to 1% in lysis buffer and treated with 50 µg/ml proteinase K (Invitrogen) for one hour at 37°C.

    Techniques: Infection, Mouse Assay, Western Blot, Migration

    Analysis of spongiform changes in C12 WT and KO brain sections (A, i and iii) show a similar amount of vacuolation, indicated by arrows, in the hippocampus of prion inoculated mice but no vacuolation in uninoculated mice [C12 WT is shown in (A, i)]. The amount of gliosis was examined by staining for GFAP, which did not show staining in uninoculated samples [C12 WT is shown in (A, iv)] but showed abundant staining in prion inoculated samples from C12 WT and C12 KO (v and vi). For all of these parameters, blinded analysis did not reveal any differences between prion inoculated C12 KO and control brains. (B) The amount of proteinase K resistant PrP was assayed in whole brain homogenates taken from prion inoculated C12 WT (n = 5) and C12 KO (n = 5) mice (treated for 50ug/ml PK for one hour at 37C), which all showed ample PK resistant PrP by immunoblotting with SAF83. Total PrP inputs are shown to ensure equivalent loading.

    Journal: Prion

    Article Title: Prion Pathogenesis is Independent of Caspase-12

    doi:

    Figure Lengend Snippet: Analysis of spongiform changes in C12 WT and KO brain sections (A, i and iii) show a similar amount of vacuolation, indicated by arrows, in the hippocampus of prion inoculated mice but no vacuolation in uninoculated mice [C12 WT is shown in (A, i)]. The amount of gliosis was examined by staining for GFAP, which did not show staining in uninoculated samples [C12 WT is shown in (A, iv)] but showed abundant staining in prion inoculated samples from C12 WT and C12 KO (v and vi). For all of these parameters, blinded analysis did not reveal any differences between prion inoculated C12 KO and control brains. (B) The amount of proteinase K resistant PrP was assayed in whole brain homogenates taken from prion inoculated C12 WT (n = 5) and C12 KO (n = 5) mice (treated for 50ug/ml PK for one hour at 37C), which all showed ample PK resistant PrP by immunoblotting with SAF83. Total PrP inputs are shown to ensure equivalent loading.

    Article Snippet: For proteinase K treatment, 10% brain homogenates from terminally ill WT or Caspase-12 KOs were diluted to 1% in lysis buffer and treated with 50 µg/ml proteinase K (Invitrogen) for one hour at 37°C.

    Techniques: Mouse Assay, Staining

    GORASP2 is localized to autophagosomes and lysosomes upon starvation. ( a ) GORASP2 colocalizes with GFP-LC3 upon starvation. GFP-LC3 HeLa cells were treated with growth medium (ctr), BafA1, EBSS or EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, GOLGA2 and DNA. The bottom row shows higher magnifications of the indicated area in the above row. Scale bar: 10 µm in the upper four rows, 3 µm in the bottom row. ( b ) Quantification of ( a ) for the percentage of GORASP2 puncta that colocalized with GFP-LC3. ( c ) GORASP2 colocalizes with LC3 and LAMP2 but not EEA1 or LAMP1 upon starvation. HeLa cells were treated with EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, LC3, EEA1, LAMP1, or LAMP2 and DNA as indicated. The three rows on the right are higher magnifications of the boxed area in the three rows on the left. Scale bars: 10 µm in the left rows, 3 µm in the right rows. ( d ) Quantification of ( c ) for the percentage of GORASP2 puncta that colocalized with LC3, EEA1, LAMP1 or LAMP2. ( e ) Western blot of the proteinase K protection assay. HeLa cells were treated with growth medium (ctr) or EBSS and 400 nM BafA1 (E + B) for 4 h, then the collected PNS were equally divided into three tubes, one left untreated, one was incubated with 2.5 μg/ml protease K (PK) only, and one was treated with both PK and 1% TritonX-100 (TX-100) for 10 min.

    Journal: Autophagy

    Article Title: GORASP2/GRASP55 collaborates with the PtdIns3K UVRAG complex to facilitate autophagosome-lysosome fusion

    doi: 10.1080/15548627.2019.1596480

    Figure Lengend Snippet: GORASP2 is localized to autophagosomes and lysosomes upon starvation. ( a ) GORASP2 colocalizes with GFP-LC3 upon starvation. GFP-LC3 HeLa cells were treated with growth medium (ctr), BafA1, EBSS or EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, GOLGA2 and DNA. The bottom row shows higher magnifications of the indicated area in the above row. Scale bar: 10 µm in the upper four rows, 3 µm in the bottom row. ( b ) Quantification of ( a ) for the percentage of GORASP2 puncta that colocalized with GFP-LC3. ( c ) GORASP2 colocalizes with LC3 and LAMP2 but not EEA1 or LAMP1 upon starvation. HeLa cells were treated with EBSS and 400 nM BafA1 (E + B) for 4 h, stained for GORASP2, LC3, EEA1, LAMP1, or LAMP2 and DNA as indicated. The three rows on the right are higher magnifications of the boxed area in the three rows on the left. Scale bars: 10 µm in the left rows, 3 µm in the right rows. ( d ) Quantification of ( c ) for the percentage of GORASP2 puncta that colocalized with LC3, EEA1, LAMP1 or LAMP2. ( e ) Western blot of the proteinase K protection assay. HeLa cells were treated with growth medium (ctr) or EBSS and 400 nM BafA1 (E + B) for 4 h, then the collected PNS were equally divided into three tubes, one left untreated, one was incubated with 2.5 μg/ml protease K (PK) only, and one was treated with both PK and 1% TritonX-100 (TX-100) for 10 min.

    Article Snippet: Each PNS was equally divided into three tubes, one was left untreated, one was incubated with 2.5 μg/ml Protease K (Thermo Fisher Scientific, AM2542), and the other was treated with both protease K and 1% Triton X-100 (from 20% stock) for 10 min on ice.

    Techniques: Staining, Western Blot, Incubation