bsa  (Thermo Fisher)


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    Structured Review

    Thermo Fisher bsa
    EV subtype-specific enrichment and characterization. (A) Schematic depiction of the isolation protocol for sEVs and m/lEVs. (B and C) Size distributions and protein concentrations of sEV- and m/lEV-enriched fractions derived from cancer (MDA-MB-231, MDA-MB-468) and non-cancerous cells (keratinocyte, fibroblast) measured by NTA and <t>Bradford</t> assay, respectively. (D) Western blot analysis of the EV marker proteins, CD81, CD63, CD9, TSG101, and flotillin-1 in sEV- and m/lEV-enriched fractions from each cell type.
    Bsa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsa/product/Thermo Fisher
    Average 88 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    bsa - by Bioz Stars, 2022-11
    88/100 stars

    Images

    1) Product Images from "Phosphatidylserine-exposing medium/large extracellular vesicles: potential cancer biomarkers"

    Article Title: Phosphatidylserine-exposing medium/large extracellular vesicles: potential cancer biomarkers

    Journal: bioRxiv

    doi: 10.1101/2022.11.17.516966

    EV subtype-specific enrichment and characterization. (A) Schematic depiction of the isolation protocol for sEVs and m/lEVs. (B and C) Size distributions and protein concentrations of sEV- and m/lEV-enriched fractions derived from cancer (MDA-MB-231, MDA-MB-468) and non-cancerous cells (keratinocyte, fibroblast) measured by NTA and Bradford assay, respectively. (D) Western blot analysis of the EV marker proteins, CD81, CD63, CD9, TSG101, and flotillin-1 in sEV- and m/lEV-enriched fractions from each cell type.
    Figure Legend Snippet: EV subtype-specific enrichment and characterization. (A) Schematic depiction of the isolation protocol for sEVs and m/lEVs. (B and C) Size distributions and protein concentrations of sEV- and m/lEV-enriched fractions derived from cancer (MDA-MB-231, MDA-MB-468) and non-cancerous cells (keratinocyte, fibroblast) measured by NTA and Bradford assay, respectively. (D) Western blot analysis of the EV marker proteins, CD81, CD63, CD9, TSG101, and flotillin-1 in sEV- and m/lEV-enriched fractions from each cell type.

    Techniques Used: Isolation, Derivative Assay, Multiple Displacement Amplification, Bradford Assay, Western Blot, Marker

    2) Product Images from "Phosphatidylserine-exposing medium/large extracellular vesicles: potential cancer biomarkers"

    Article Title: Phosphatidylserine-exposing medium/large extracellular vesicles: potential cancer biomarkers

    Journal: bioRxiv

    doi: 10.1101/2022.11.17.516966

    EV subtype-specific enrichment and characterization. (A) Schematic depiction of the isolation protocol for sEVs and m/lEVs. (B and C) Size distributions and protein concentrations of sEV- and m/lEV-enriched fractions derived from cancer (MDA-MB-231, MDA-MB-468) and non-cancerous cells (keratinocyte, fibroblast) measured by NTA and Bradford assay, respectively. (D) Western blot analysis of the EV marker proteins, CD81, CD63, CD9, TSG101, and flotillin-1 in sEV- and m/lEV-enriched fractions from each cell type.
    Figure Legend Snippet: EV subtype-specific enrichment and characterization. (A) Schematic depiction of the isolation protocol for sEVs and m/lEVs. (B and C) Size distributions and protein concentrations of sEV- and m/lEV-enriched fractions derived from cancer (MDA-MB-231, MDA-MB-468) and non-cancerous cells (keratinocyte, fibroblast) measured by NTA and Bradford assay, respectively. (D) Western blot analysis of the EV marker proteins, CD81, CD63, CD9, TSG101, and flotillin-1 in sEV- and m/lEV-enriched fractions from each cell type.

    Techniques Used: Isolation, Derivative Assay, Multiple Displacement Amplification, Bradford Assay, Western Blot, Marker

    3) Product Images from "Computationally designed GPCR quaternary structures bias signaling pathway activation"

    Article Title: Computationally designed GPCR quaternary structures bias signaling pathway activation

    Journal: Nature Communications

    doi: 10.1038/s41467-022-34382-7

    Computational design of CXCR4 associations with specific conformations. a Mutations designed to selectively stabilize the CXCR4 open-dimer conformation without affecting CXCR4 monomer stability were identified in the extracellular and TMH regions. b Mutations designed to selectively stabilize the CXCR4 closed-dimer conformation without affecting CXCR4 monomer stability were identified in the extracellular region. Key atomic contacts are represented as red dotted lines. c Schematic conformational energy landscapes of CXCR4 dimerization in the inactive and active states for the open-dimer stabilizing designs. d Schematic conformational energy landscapes of CXCR4 dimerization in the inactive and active states for the closed-dimer stabilizing designs. c , d The dimerization energies reported in Supplementary Table 1 were used to plot the energy landscapes. The monomer energies and energy barriers between states are fictitious and were not predicted by our simulations. e Ranking of the CXCR4 variants based on changes in buried surface area upon dimerization (ΔSASA) calculated from the predicted models in the active state. The ΔSASA is reported for the most occupied dimer conformation for each variant: L194R open-dimer state, WT open-dimer state, N192W closed-dimer state, W195L closed-dimer state. Larger buried ΔSASA are predicted to correlate with enhanced dimerization propensity (see Supplementary Table 2 ).
    Figure Legend Snippet: Computational design of CXCR4 associations with specific conformations. a Mutations designed to selectively stabilize the CXCR4 open-dimer conformation without affecting CXCR4 monomer stability were identified in the extracellular and TMH regions. b Mutations designed to selectively stabilize the CXCR4 closed-dimer conformation without affecting CXCR4 monomer stability were identified in the extracellular region. Key atomic contacts are represented as red dotted lines. c Schematic conformational energy landscapes of CXCR4 dimerization in the inactive and active states for the open-dimer stabilizing designs. d Schematic conformational energy landscapes of CXCR4 dimerization in the inactive and active states for the closed-dimer stabilizing designs. c , d The dimerization energies reported in Supplementary Table 1 were used to plot the energy landscapes. The monomer energies and energy barriers between states are fictitious and were not predicted by our simulations. e Ranking of the CXCR4 variants based on changes in buried surface area upon dimerization (ΔSASA) calculated from the predicted models in the active state. The ΔSASA is reported for the most occupied dimer conformation for each variant: L194R open-dimer state, WT open-dimer state, N192W closed-dimer state, W195L closed-dimer state. Larger buried ΔSASA are predicted to correlate with enhanced dimerization propensity (see Supplementary Table 2 ).

    Techniques Used: Variant Assay

    4) Product Images from "Protein and RNA ADP-ribosylation detection is influenced by sample preparation and reagents used"

    Article Title: Protein and RNA ADP-ribosylation detection is influenced by sample preparation and reagents used

    Journal: Life Science Alliance

    doi: 10.26508/lsa.202201455

    Tests with additional ADPr antibodies HCA354 and HCA355. (A) In vitro ADP-ribosylation reactions were performed with indicated transferases and analysed using Western blot. (B) Peptides with ADPr linked to different amino acids were slot-blotted and analysed using indicated antibodies. (C) AMPylated proteins and their non-modified controls were slot-blotted and analysed using the indicated antibodies. Automodified PARP10cat serves as a positive control. (D) Differentially capped RNAs were slot-blotted and analysed using the indicated antibodies. Automodified PARP10cat serves as a positive control. (E) NAD+, NADH, and ADPr were cross-linked to BSA using PFA and slot-blotted. PARP10cat and automodified PARP10cat serve as a negative and a positive control, respectively.
    Figure Legend Snippet: Tests with additional ADPr antibodies HCA354 and HCA355. (A) In vitro ADP-ribosylation reactions were performed with indicated transferases and analysed using Western blot. (B) Peptides with ADPr linked to different amino acids were slot-blotted and analysed using indicated antibodies. (C) AMPylated proteins and their non-modified controls were slot-blotted and analysed using the indicated antibodies. Automodified PARP10cat serves as a positive control. (D) Differentially capped RNAs were slot-blotted and analysed using the indicated antibodies. Automodified PARP10cat serves as a positive control. (E) NAD+, NADH, and ADPr were cross-linked to BSA using PFA and slot-blotted. PARP10cat and automodified PARP10cat serve as a negative and a positive control, respectively.

    Techniques Used: In Vitro, Western Blot, Modification, Positive Control

    ADP-ribosylation detection reagents stain different structures depending on the fixation method used. (A) HeLa cells were seeded onto glass coverslips and fixed using PFA, ice-cold methanol, or glyoxal. Reagent V was used to stain the cells, and DAPI was applied to stain the nuclei. (B) As in (A), but before fixation cells were treated with 0.5 mM H 2 O 2 for 10 min. All images were taken using a confocal microscope with identical laser intensity and settings across all samples. (C) HeLa cells were seeded as in (A) and fixed using PFA. Fixed samples were treated with either DNase or RNase, HgCl 2 , or hydroxylamine. ADP-ribosylation was visualised using Reagent V, and images were taken using confocal microscopy at the same settings for each sample. (D) Indicated metabolites were cross-linked to BSA using PFA and slot-blotted. Blots were incubated with the indicated antibodies and detected using chemiluminescence. Scale bars in (A, B, C) represent 10 µ M.
    Figure Legend Snippet: ADP-ribosylation detection reagents stain different structures depending on the fixation method used. (A) HeLa cells were seeded onto glass coverslips and fixed using PFA, ice-cold methanol, or glyoxal. Reagent V was used to stain the cells, and DAPI was applied to stain the nuclei. (B) As in (A), but before fixation cells were treated with 0.5 mM H 2 O 2 for 10 min. All images were taken using a confocal microscope with identical laser intensity and settings across all samples. (C) HeLa cells were seeded as in (A) and fixed using PFA. Fixed samples were treated with either DNase or RNase, HgCl 2 , or hydroxylamine. ADP-ribosylation was visualised using Reagent V, and images were taken using confocal microscopy at the same settings for each sample. (D) Indicated metabolites were cross-linked to BSA using PFA and slot-blotted. Blots were incubated with the indicated antibodies and detected using chemiluminescence. Scale bars in (A, B, C) represent 10 µ M.

    Techniques Used: Staining, Microscopy, Confocal Microscopy, Incubation

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    Thermo Fisher bodipy fl c5 ceramide complexed to bsa
    Bodipy Fl C5 Ceramide Complexed To Bsa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bsa fraction v
    Bsa Fraction V, supplied by Thermo Fisher, 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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    Thermo Fisher dq green bsa
    Leucine independence is a feature of naïve pluripotency. ( A and B ) <t>DQ-BSA</t> quantification in Rex1-GFP (A) and Nanog-GFP (B) S/L cultured ESCs. ( C ) Nanog-GFP expression in S/L cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( D ) RT-qPCR of pluripotency-associated ( Esrrb, Klf4, Nanog, Zfp42/Rex1, Pou5f1/Oct4 ) and early differentiation associated genes ( Fgf5, Oct6, Otx2, T ) in S/L-cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( E ) Quantification of differentiated, mixed and undifferentiated colonies formed by ESCs seeded at clonal density in S/L medium following 24 h culture in leucine-deprived (−Leu) or control (+Leu) conditions. Data are mean ± SD, n=5 independent samples (A and B) or n=3 independent samples (D and E). Significance was assessed by unpaired two-tailed t test [(A), (B), and (E)] (** p
    Dq Green Bsa, supplied by Thermo Fisher, 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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    Thermo Fisher bsa
    Leucine independence is a feature of naïve pluripotency. ( A and B ) <t>DQ-BSA</t> quantification in Rex1-GFP (A) and Nanog-GFP (B) S/L cultured ESCs. ( C ) Nanog-GFP expression in S/L cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( D ) RT-qPCR of pluripotency-associated ( Esrrb, Klf4, Nanog, Zfp42/Rex1, Pou5f1/Oct4 ) and early differentiation associated genes ( Fgf5, Oct6, Otx2, T ) in S/L-cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( E ) Quantification of differentiated, mixed and undifferentiated colonies formed by ESCs seeded at clonal density in S/L medium following 24 h culture in leucine-deprived (−Leu) or control (+Leu) conditions. Data are mean ± SD, n=5 independent samples (A and B) or n=3 independent samples (D and E). Significance was assessed by unpaired two-tailed t test [(A), (B), and (E)] (** p
    Bsa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsa/product/Thermo Fisher
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsa - by Bioz Stars, 2022-11
    88/100 stars
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    Image Search Results


    Leucine independence is a feature of naïve pluripotency. ( A and B ) DQ-BSA quantification in Rex1-GFP (A) and Nanog-GFP (B) S/L cultured ESCs. ( C ) Nanog-GFP expression in S/L cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( D ) RT-qPCR of pluripotency-associated ( Esrrb, Klf4, Nanog, Zfp42/Rex1, Pou5f1/Oct4 ) and early differentiation associated genes ( Fgf5, Oct6, Otx2, T ) in S/L-cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( E ) Quantification of differentiated, mixed and undifferentiated colonies formed by ESCs seeded at clonal density in S/L medium following 24 h culture in leucine-deprived (−Leu) or control (+Leu) conditions. Data are mean ± SD, n=5 independent samples (A and B) or n=3 independent samples (D and E). Significance was assessed by unpaired two-tailed t test [(A), (B), and (E)] (** p

    Journal: bioRxiv

    Article Title: Amino acid intake strategies define pluripotent cell states

    doi: 10.1101/2022.11.16.516803

    Figure Lengend Snippet: Leucine independence is a feature of naïve pluripotency. ( A and B ) DQ-BSA quantification in Rex1-GFP (A) and Nanog-GFP (B) S/L cultured ESCs. ( C ) Nanog-GFP expression in S/L cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( D ) RT-qPCR of pluripotency-associated ( Esrrb, Klf4, Nanog, Zfp42/Rex1, Pou5f1/Oct4 ) and early differentiation associated genes ( Fgf5, Oct6, Otx2, T ) in S/L-cultured ESCs subjected to 24 h of leucine (Leu) replete or depleted conditions. ( E ) Quantification of differentiated, mixed and undifferentiated colonies formed by ESCs seeded at clonal density in S/L medium following 24 h culture in leucine-deprived (−Leu) or control (+Leu) conditions. Data are mean ± SD, n=5 independent samples (A and B) or n=3 independent samples (D and E). Significance was assessed by unpaired two-tailed t test [(A), (B), and (E)] (** p

    Article Snippet: Cells were allowed to grow for additional 24 h before treatment with 100 µg/mL DQ Green BSA (Thermo Fisher Scientific) for 1 h. Cells were then washed twice in PBS and imaged using a Zeiss Axio Vert.A1 inverted microscope with a black and white camera (Axiocam MRm).

    Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Two Tailed Test

    Lysosomal metabolism facilitates amino acid independence. ( A and B ) Quantification of LysoTracker signal in S/L and S/L+2i cultured ESCs (A) and Nanog-GFP S/L cultured ESCs (B). ( C and D ) LysoTracker (C) and DQ-BSA (D) signal in Tfe3-ERT2-expressing S/L cultured ESCs treated with vehicle (ethanol, EtOH) or tamoxifen (Tam). ( E ) Population doublings of Tfe3-ERT2-expressing S/L cultured ESCs treated with EtOH or Tam and subjected to leucine (Leu) withdrawal for 48 h. ( F ) Schematic of experimental setup to induce exit from naïve pluripotency. ( G ) Expression of KEGG lysosome genes in ESCs following 2i/L withdrawal for the indicated times. ( H and I ) Quantification of LysoTracker (H) and DQ-BSA (I) signal in 2i/L cultured ESCs subjected to 2i/L withdrawal for 12, 24 and 40 h. ( J and K ) Alkaline phosphatase (AP) staining of colony formation assays of S/L+2i cultured ESCs subjected to 2i/L withdrawal for 40 h in the presence or absence of leucine prior to reseeding in leucine-replete 2i/L medium to assess exit from naïve pluripotency. Representative wells are shown in J, quantification of biological replicates is shown in K. ( L ) Expression of genes encoding plasma membrane amino acid transporters in ESCs following 2i/L withdrawal for the indicated times. ( M ) Volcano plot of amino acid uptake from culture media by 2i/L cultured ESCs subjected to control or -2i/L conditions for 40 h. Media incubated with cells for the last 24 h of the experiment was assayed. Data are mean ± SD, n=3 independent samples (A, C-E, H, I, K), n=5 independent samples (B), n=2 independent samples (G and L), n=6 independent samples (M). Significance was assessed by unpaired two-tailed t test [(A) to (E), (K)] (** p

    Journal: bioRxiv

    Article Title: Amino acid intake strategies define pluripotent cell states

    doi: 10.1101/2022.11.16.516803

    Figure Lengend Snippet: Lysosomal metabolism facilitates amino acid independence. ( A and B ) Quantification of LysoTracker signal in S/L and S/L+2i cultured ESCs (A) and Nanog-GFP S/L cultured ESCs (B). ( C and D ) LysoTracker (C) and DQ-BSA (D) signal in Tfe3-ERT2-expressing S/L cultured ESCs treated with vehicle (ethanol, EtOH) or tamoxifen (Tam). ( E ) Population doublings of Tfe3-ERT2-expressing S/L cultured ESCs treated with EtOH or Tam and subjected to leucine (Leu) withdrawal for 48 h. ( F ) Schematic of experimental setup to induce exit from naïve pluripotency. ( G ) Expression of KEGG lysosome genes in ESCs following 2i/L withdrawal for the indicated times. ( H and I ) Quantification of LysoTracker (H) and DQ-BSA (I) signal in 2i/L cultured ESCs subjected to 2i/L withdrawal for 12, 24 and 40 h. ( J and K ) Alkaline phosphatase (AP) staining of colony formation assays of S/L+2i cultured ESCs subjected to 2i/L withdrawal for 40 h in the presence or absence of leucine prior to reseeding in leucine-replete 2i/L medium to assess exit from naïve pluripotency. Representative wells are shown in J, quantification of biological replicates is shown in K. ( L ) Expression of genes encoding plasma membrane amino acid transporters in ESCs following 2i/L withdrawal for the indicated times. ( M ) Volcano plot of amino acid uptake from culture media by 2i/L cultured ESCs subjected to control or -2i/L conditions for 40 h. Media incubated with cells for the last 24 h of the experiment was assayed. Data are mean ± SD, n=3 independent samples (A, C-E, H, I, K), n=5 independent samples (B), n=2 independent samples (G and L), n=6 independent samples (M). Significance was assessed by unpaired two-tailed t test [(A) to (E), (K)] (** p

    Article Snippet: Cells were allowed to grow for additional 24 h before treatment with 100 µg/mL DQ Green BSA (Thermo Fisher Scientific) for 1 h. Cells were then washed twice in PBS and imaged using a Zeiss Axio Vert.A1 inverted microscope with a black and white camera (Axiocam MRm).

    Techniques: Cell Culture, Expressing, Staining, Incubation, Two Tailed Test

    Digestive amino acid metabolism is attenuated with developmental progression. ( A and B ) Violin plots of scRNA-seq data 40 showing the expression distribution of KEGG lysosome (A) and plasma amino acid transporter (B) genes in the E3.5 ICM and E4.5 and E5.5 epiblast. ( C ) Gene set enrichment analysis of 2i/L cultured ESCs and EpiLCs 43 showing enrichment of KEGG Lysosome genes among genes upregulated in 2i/L and plasma membrane amino acid transporter genes enriched in the EpiLC state( C ) Gene set enrichment analysis of 2i/L cultured ESCs and EpiLCs 43 showing enrichment of KEGG Lysosome genes among genes upregulated in 2i/L and plasma membrane amino acid transporter genes enriched in genes upregulated in the EpiLC state. ( D and E ) Quantification of LysoTracker (D) and DQ-BSA (E) in S/L+2i cultured ESCs and EpiLCs. Data are mean ± SD, n=3 independent samples (D and E). Significance was assessed in comparison to E3.5 cells by one-way ANOVA with Sidak’s multiple comparisons post-test (**** p

    Journal: bioRxiv

    Article Title: Amino acid intake strategies define pluripotent cell states

    doi: 10.1101/2022.11.16.516803

    Figure Lengend Snippet: Digestive amino acid metabolism is attenuated with developmental progression. ( A and B ) Violin plots of scRNA-seq data 40 showing the expression distribution of KEGG lysosome (A) and plasma amino acid transporter (B) genes in the E3.5 ICM and E4.5 and E5.5 epiblast. ( C ) Gene set enrichment analysis of 2i/L cultured ESCs and EpiLCs 43 showing enrichment of KEGG Lysosome genes among genes upregulated in 2i/L and plasma membrane amino acid transporter genes enriched in the EpiLC state( C ) Gene set enrichment analysis of 2i/L cultured ESCs and EpiLCs 43 showing enrichment of KEGG Lysosome genes among genes upregulated in 2i/L and plasma membrane amino acid transporter genes enriched in genes upregulated in the EpiLC state. ( D and E ) Quantification of LysoTracker (D) and DQ-BSA (E) in S/L+2i cultured ESCs and EpiLCs. Data are mean ± SD, n=3 independent samples (D and E). Significance was assessed in comparison to E3.5 cells by one-way ANOVA with Sidak’s multiple comparisons post-test (**** p

    Article Snippet: Cells were allowed to grow for additional 24 h before treatment with 100 µg/mL DQ Green BSA (Thermo Fisher Scientific) for 1 h. Cells were then washed twice in PBS and imaged using a Zeiss Axio Vert.A1 inverted microscope with a black and white camera (Axiocam MRm).

    Techniques: Expressing, Cell Culture

    Cells in pre-implantation blastocyst engage in macropinocytosis. ( A ) Schematic of experimental workflow in E3.75 mouse blastocysts. ( B ) Representative fluorescent and differential interference contrast (DIC) images of control and EIPA pre-treated E3.75 mouse blastocysts incubated with DQ-BSA. ( C ) Representative fluorescent and DIC images of control and EIPA pre-treated ICM dissociated from E3.75 Pdgfra H2B-GFP/+ blastocysts and incubated with DQ-BSA. GFP demarcates primitive endoderm. Scale bars, 25 µm.

    Journal: bioRxiv

    Article Title: Amino acid intake strategies define pluripotent cell states

    doi: 10.1101/2022.11.16.516803

    Figure Lengend Snippet: Cells in pre-implantation blastocyst engage in macropinocytosis. ( A ) Schematic of experimental workflow in E3.75 mouse blastocysts. ( B ) Representative fluorescent and differential interference contrast (DIC) images of control and EIPA pre-treated E3.75 mouse blastocysts incubated with DQ-BSA. ( C ) Representative fluorescent and DIC images of control and EIPA pre-treated ICM dissociated from E3.75 Pdgfra H2B-GFP/+ blastocysts and incubated with DQ-BSA. GFP demarcates primitive endoderm. Scale bars, 25 µm.

    Article Snippet: Cells were allowed to grow for additional 24 h before treatment with 100 µg/mL DQ Green BSA (Thermo Fisher Scientific) for 1 h. Cells were then washed twice in PBS and imaged using a Zeiss Axio Vert.A1 inverted microscope with a black and white camera (Axiocam MRm).

    Techniques: Incubation

    ESCs engage in constitutive macropinocytosis. ( A ) Representative fluorescent and differential interference contrast (DIC) images of S/L and S/L+2i cultured ESCs treated with DQ-BSA for 1 h. ( B ) Quantification of DQ-BSA in S/L and S/L+2i cultured ESCs. Six fields were assessed per culture condition. ( C ) Flow cytometry quantification of DQ-BSA signal in S/L and S/L+2i cultured ESCs. ( D and E ) Rate of Dextran (D) and DQ-BSA (E) in S/L and S/L+2i cultured ESCs. ( F - I ) Flow cytometry quantification of DQ-BSA signal in S/L and S/L+2i cultured ESCs following pre-treatment with the indicated inhibitors. Data are mean ± SD, n=3 independent samples (C to I). Significance was assessed by unpaired two-tailed t test [(C)], (**** p

    Journal: bioRxiv

    Article Title: Amino acid intake strategies define pluripotent cell states

    doi: 10.1101/2022.11.16.516803

    Figure Lengend Snippet: ESCs engage in constitutive macropinocytosis. ( A ) Representative fluorescent and differential interference contrast (DIC) images of S/L and S/L+2i cultured ESCs treated with DQ-BSA for 1 h. ( B ) Quantification of DQ-BSA in S/L and S/L+2i cultured ESCs. Six fields were assessed per culture condition. ( C ) Flow cytometry quantification of DQ-BSA signal in S/L and S/L+2i cultured ESCs. ( D and E ) Rate of Dextran (D) and DQ-BSA (E) in S/L and S/L+2i cultured ESCs. ( F - I ) Flow cytometry quantification of DQ-BSA signal in S/L and S/L+2i cultured ESCs following pre-treatment with the indicated inhibitors. Data are mean ± SD, n=3 independent samples (C to I). Significance was assessed by unpaired two-tailed t test [(C)], (**** p

    Article Snippet: Cells were allowed to grow for additional 24 h before treatment with 100 µg/mL DQ Green BSA (Thermo Fisher Scientific) for 1 h. Cells were then washed twice in PBS and imaged using a Zeiss Axio Vert.A1 inverted microscope with a black and white camera (Axiocam MRm).

    Techniques: Cell Culture, Flow Cytometry, Two Tailed Test