ta1 Search Results


fasciculin  (Alomone Labs)
93
Alomone Labs fasciculin
Fasciculin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/fasciculin/product/Alomone Labs
Average 93 stars, based on 1 article reviews
fasciculin - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
lat1 slc7a5  (Alomone Labs)
92
Alomone Labs lat1 slc7a5
Basal expression of <t>SLC7A5/SLC3A2</t> in the erythroid lineage (A) The panel represents the flow cytometry analysis showing the different splenic cell populations in basal conditions based on the surface expression of CD71 and Ter119 in control mice. SLC3A2 expression is also represented by a color code: yellow-orange representing the strongest expression and dark green-blue the weakest or absence of expression. (B) Histograms showing SLC3A2 expression in the different splenic CD71 HI /Ter119 NEG/LOW , CD71 HI /Ter119 HI , CD71 NEG /Ter119 HI and CD71 NEG/LOW /Ter119 NEG cell populations in control (in blue) and ErGFPcre Slc7a5 LoxP / LoxP mice (in green). The IgG control (in gray) is included. A representative experiment out of 23 is shown.
Lat1 Slc7a5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/lat1 slc7a5/product/Alomone Labs
Average 92 stars, based on 1 article reviews
lat1 slc7a5 - by Bioz Stars, 2026-02
92/100 stars
  Buy from Supplier
achr inhibitor fasciculin ii  (Alomone Labs)
90
Alomone Labs achr inhibitor fasciculin ii
Basal expression of <t>SLC7A5/SLC3A2</t> in the erythroid lineage (A) The panel represents the flow cytometry analysis showing the different splenic cell populations in basal conditions based on the surface expression of CD71 and Ter119 in control mice. SLC3A2 expression is also represented by a color code: yellow-orange representing the strongest expression and dark green-blue the weakest or absence of expression. (B) Histograms showing SLC3A2 expression in the different splenic CD71 HI /Ter119 NEG/LOW , CD71 HI /Ter119 HI , CD71 NEG /Ter119 HI and CD71 NEG/LOW /Ter119 NEG cell populations in control (in blue) and ErGFPcre Slc7a5 LoxP / LoxP mice (in green). The IgG control (in gray) is included. A representative experiment out of 23 is shown.
Achr Inhibitor Fasciculin Ii, 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
https://www.bioz.com/result/achr inhibitor fasciculin ii/product/Alomone Labs
Average 90 stars, based on 1 article reviews
achr inhibitor fasciculin ii - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
pcmv6 slc7a5  (OriGene)
93
OriGene pcmv6 slc7a5
( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of <t>Slc7a5</t> mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.
Pcmv6 Slc7a5, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pcmv6 slc7a5/product/OriGene
Average 93 stars, based on 1 article reviews
pcmv6 slc7a5 - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
zadaxin® brand ta1  (SciClone Inc)
90
SciClone Inc zadaxin® brand ta1
( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of <t>Slc7a5</t> mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.
Zadaxin® Brand Ta1, supplied by SciClone Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/zadaxin® brand ta1/product/SciClone Inc
Average 90 stars, based on 1 article reviews
zadaxin® brand ta1 - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
gbsa(chi/ta) 1 nps  (CH Instruments)
90
CH Instruments gbsa(chi/ta) 1 nps
( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of <t>Slc7a5</t> mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.
Gbsa(Chi/Ta) 1 Nps, supplied by CH Instruments, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gbsa(chi/ta) 1 nps/product/CH Instruments
Average 90 stars, based on 1 article reviews
gbsa(chi/ta) 1 nps - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
titanium nail ta1  (Huachuang Securities Co Ltd)
90
Huachuang Securities Co Ltd titanium nail ta1
( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of <t>Slc7a5</t> mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.
Titanium Nail Ta1, supplied by Huachuang Securities Co Ltd, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/titanium nail ta1/product/Huachuang Securities Co Ltd
Average 90 stars, based on 1 article reviews
titanium nail ta1 - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
ta1 texture analyzer  (Lloyd Instruments)
90
Lloyd Instruments ta1 texture analyzer
( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of <t>Slc7a5</t> mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.
Ta1 Texture Analyzer, supplied by Lloyd Instruments, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ta1 texture analyzer/product/Lloyd Instruments
Average 90 stars, based on 1 article reviews
ta1 texture analyzer - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
ta-1  (SciClone Inc)
90
SciClone Inc ta-1
( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of <t>Slc7a5</t> mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.
Ta 1, supplied by SciClone Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ta-1/product/SciClone Inc
Average 90 stars, based on 1 article reviews
ta-1 - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
cd44 aptamer ta1  (Eurofins)
90
Eurofins cd44 aptamer ta1
Schematic illustration of the work which showed biologically transformable NPs <t>(aptamer</t> and peptide-modified DTX-loaded pH and glutathione (GSH) dual responsive polymeric micelles) with altered performance after exposure to overexpressed MMP-9 in breast cancer TME. The synthesized NPs generally exhibit relatively neutral biological activity in the bloodstream and show minimal interaction with blood proteins (especially opsonins), as noted in the earlier article. However, upon entering the tumor microenvironment, and following the breakdown of the fusogenic peptide, the hidden aptamers <t>(TA1)</t> between these peptides, along with the remaining peptide sequence (SRL-2), suddenly become exposed. These aptamers alongside of SRL-2 peptide (10 amino acid remaining after cleavage of fusogenic peptide) then synergistically recognize the <t>CD44</t> and LRP-1, respectively on cancer cells, demonstrating a significantly high cellular uptake efficiency. Essentially, our initial nanoparticles function like a shuttle, as depicted, that upon reaching the tumor boundary (illustrated by the gray bubble around the tumor in the schematic), loses part of its structure. The remaining part of the structure, resembling a missile cap with dual targeting ends, identifies the target cell receptors and effectively delivers the chemotherapeutic drug with high efficacy (at 1/10th of the therapeutic concentration) to the cancer cells.
Cd44 Aptamer Ta1, supplied by Eurofins, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/cd44 aptamer ta1/product/Eurofins
Average 90 stars, based on 1 article reviews
cd44 aptamer ta1 - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
ethoxylated (20) trimethylolpropane triacrylate (peg-ta) 1.2k  (Sartomer USA LLC)
90
Sartomer USA LLC ethoxylated (20) trimethylolpropane triacrylate (peg-ta) 1.2k
Schematic illustration of the work which showed biologically transformable NPs <t>(aptamer</t> and peptide-modified DTX-loaded pH and glutathione (GSH) dual responsive polymeric micelles) with altered performance after exposure to overexpressed MMP-9 in breast cancer TME. The synthesized NPs generally exhibit relatively neutral biological activity in the bloodstream and show minimal interaction with blood proteins (especially opsonins), as noted in the earlier article. However, upon entering the tumor microenvironment, and following the breakdown of the fusogenic peptide, the hidden aptamers <t>(TA1)</t> between these peptides, along with the remaining peptide sequence (SRL-2), suddenly become exposed. These aptamers alongside of SRL-2 peptide (10 amino acid remaining after cleavage of fusogenic peptide) then synergistically recognize the <t>CD44</t> and LRP-1, respectively on cancer cells, demonstrating a significantly high cellular uptake efficiency. Essentially, our initial nanoparticles function like a shuttle, as depicted, that upon reaching the tumor boundary (illustrated by the gray bubble around the tumor in the schematic), loses part of its structure. The remaining part of the structure, resembling a missile cap with dual targeting ends, identifies the target cell receptors and effectively delivers the chemotherapeutic drug with high efficacy (at 1/10th of the therapeutic concentration) to the cancer cells.
Ethoxylated (20) Trimethylolpropane Triacrylate (Peg Ta) 1.2k, supplied by Sartomer USA LLC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ethoxylated (20) trimethylolpropane triacrylate (peg-ta) 1.2k/product/Sartomer USA LLC
Average 90 stars, based on 1 article reviews
ethoxylated (20) trimethylolpropane triacrylate (peg-ta) 1.2k - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier
ta1-ko mice  (Charles River Laboratories)
90
Charles River Laboratories ta1-ko mice
Schematic illustration of the work which showed biologically transformable NPs <t>(aptamer</t> and peptide-modified DTX-loaded pH and glutathione (GSH) dual responsive polymeric micelles) with altered performance after exposure to overexpressed MMP-9 in breast cancer TME. The synthesized NPs generally exhibit relatively neutral biological activity in the bloodstream and show minimal interaction with blood proteins (especially opsonins), as noted in the earlier article. However, upon entering the tumor microenvironment, and following the breakdown of the fusogenic peptide, the hidden aptamers <t>(TA1)</t> between these peptides, along with the remaining peptide sequence (SRL-2), suddenly become exposed. These aptamers alongside of SRL-2 peptide (10 amino acid remaining after cleavage of fusogenic peptide) then synergistically recognize the <t>CD44</t> and LRP-1, respectively on cancer cells, demonstrating a significantly high cellular uptake efficiency. Essentially, our initial nanoparticles function like a shuttle, as depicted, that upon reaching the tumor boundary (illustrated by the gray bubble around the tumor in the schematic), loses part of its structure. The remaining part of the structure, resembling a missile cap with dual targeting ends, identifies the target cell receptors and effectively delivers the chemotherapeutic drug with high efficacy (at 1/10th of the therapeutic concentration) to the cancer cells.
Ta1 Ko Mice, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ta1-ko mice/product/Charles River Laboratories
Average 90 stars, based on 1 article reviews
ta1-ko mice - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier

Image Search Results


Basal expression of SLC7A5/SLC3A2 in the erythroid lineage (A) The panel represents the flow cytometry analysis showing the different splenic cell populations in basal conditions based on the surface expression of CD71 and Ter119 in control mice. SLC3A2 expression is also represented by a color code: yellow-orange representing the strongest expression and dark green-blue the weakest or absence of expression. (B) Histograms showing SLC3A2 expression in the different splenic CD71 HI /Ter119 NEG/LOW , CD71 HI /Ter119 HI , CD71 NEG /Ter119 HI and CD71 NEG/LOW /Ter119 NEG cell populations in control (in blue) and ErGFPcre Slc7a5 LoxP / LoxP mice (in green). The IgG control (in gray) is included. A representative experiment out of 23 is shown.

Journal: iScience

Article Title: Erythroid SLC7A5/SLC3A2 amino acid carrier controls red blood cell size and maturation

doi: 10.1016/j.isci.2022.105739

Figure Lengend Snippet: Basal expression of SLC7A5/SLC3A2 in the erythroid lineage (A) The panel represents the flow cytometry analysis showing the different splenic cell populations in basal conditions based on the surface expression of CD71 and Ter119 in control mice. SLC3A2 expression is also represented by a color code: yellow-orange representing the strongest expression and dark green-blue the weakest or absence of expression. (B) Histograms showing SLC3A2 expression in the different splenic CD71 HI /Ter119 NEG/LOW , CD71 HI /Ter119 HI , CD71 NEG /Ter119 HI and CD71 NEG/LOW /Ter119 NEG cell populations in control (in blue) and ErGFPcre Slc7a5 LoxP / LoxP mice (in green). The IgG control (in gray) is included. A representative experiment out of 23 is shown.

Article Snippet: The membranes were then blocked and probed with antibodies against: LAT1 (SLC7A5) (ANT-105, Alomone Labs, Jerusalem, Israel); β-actin (A3854, Sigma-Aldrich).

Techniques: Expressing, Flow Cytometry, Control

SLC7A5/SLC3A2 expression in splenic erythroblasts during erythropoietic stress (A) Representative flow cytometry analysis showing the different splenic cell populations in control and PHZ-treated mice based on the surface expression of CD71 and Ter119: CD71 HI /Ter119 NEG/LOW , CD71 HI /Ter119 HI , and CD71 NEG/LOW /Ter119 NEG . (B) Relative Slc7a5 mRNA expression in the spleen of control (n = 8) and ErGFPcre Slc7a5 LoxP / LoxP (n = 3) mice in baseline conditions as well as control (n = 6) and ErGFPcre Slc7a5 LoxP / LoxP (n = 5) upon PHZ treatment. Data are shown as mean ± SEM. Statistical analysis was performed using a two-tailed Student’s t test with Welch’s correction to compare the groups as indicated in the figure (∗p < 0.05). (C) Histograms showing SLC3A2 expression in each of the populations indicated in untreated control mice (in blue) and control mice treated with PHZ (in red) are shown in left panels. In the right panels, the same histograms of the left panels are shown, but overlaying the histogram of SLC3A2 expression of ErGFPcre Slc7a5 LoxP/LoxP (in green) mice treated with PHZ. The IgG-APC control is presented in gray. For A and C a representative experiment out of 9 is shown. (D) Representative flow cytometry analysis of DRAQ5 expression versus forward scatter (FSC) of the splenic CD71 HI /Ter119 HI erythroid population of PHZ-treated control mice. Two CD71 HI /Ter119 HI populations differing in DRAQ5 signal can be distinguished, DRAQ5 HI and DRAQ5 LOW . The dashed line represents unlabeled cells. (E) Representative histograms of SLC3A2 expression in CD71 HI /Ter119 HI /DRAQ5 HI and CD71 HI /Ter119 HI /DRAQ5 LOW populations in control mice treated with PHZ (in red) and ErGFPcre Slc7a5 LoxP/LoxP (in green) mice treated with PHZ are shown. The negative signal of the IgG-PyC control is shown in gray. For D and E a representative experiment out of 7 is shown.

Journal: iScience

Article Title: Erythroid SLC7A5/SLC3A2 amino acid carrier controls red blood cell size and maturation

doi: 10.1016/j.isci.2022.105739

Figure Lengend Snippet: SLC7A5/SLC3A2 expression in splenic erythroblasts during erythropoietic stress (A) Representative flow cytometry analysis showing the different splenic cell populations in control and PHZ-treated mice based on the surface expression of CD71 and Ter119: CD71 HI /Ter119 NEG/LOW , CD71 HI /Ter119 HI , and CD71 NEG/LOW /Ter119 NEG . (B) Relative Slc7a5 mRNA expression in the spleen of control (n = 8) and ErGFPcre Slc7a5 LoxP / LoxP (n = 3) mice in baseline conditions as well as control (n = 6) and ErGFPcre Slc7a5 LoxP / LoxP (n = 5) upon PHZ treatment. Data are shown as mean ± SEM. Statistical analysis was performed using a two-tailed Student’s t test with Welch’s correction to compare the groups as indicated in the figure (∗p < 0.05). (C) Histograms showing SLC3A2 expression in each of the populations indicated in untreated control mice (in blue) and control mice treated with PHZ (in red) are shown in left panels. In the right panels, the same histograms of the left panels are shown, but overlaying the histogram of SLC3A2 expression of ErGFPcre Slc7a5 LoxP/LoxP (in green) mice treated with PHZ. The IgG-APC control is presented in gray. For A and C a representative experiment out of 9 is shown. (D) Representative flow cytometry analysis of DRAQ5 expression versus forward scatter (FSC) of the splenic CD71 HI /Ter119 HI erythroid population of PHZ-treated control mice. Two CD71 HI /Ter119 HI populations differing in DRAQ5 signal can be distinguished, DRAQ5 HI and DRAQ5 LOW . The dashed line represents unlabeled cells. (E) Representative histograms of SLC3A2 expression in CD71 HI /Ter119 HI /DRAQ5 HI and CD71 HI /Ter119 HI /DRAQ5 LOW populations in control mice treated with PHZ (in red) and ErGFPcre Slc7a5 LoxP/LoxP (in green) mice treated with PHZ are shown. The negative signal of the IgG-PyC control is shown in gray. For D and E a representative experiment out of 7 is shown.

Article Snippet: The membranes were then blocked and probed with antibodies against: LAT1 (SLC7A5) (ANT-105, Alomone Labs, Jerusalem, Israel); β-actin (A3854, Sigma-Aldrich).

Techniques: Expressing, Flow Cytometry, Control, Two Tailed Test

SLC7A5/SLC3A2 expression in circulating CD71 HI /Ter119 HI reticulocytes under anemic conditions and in response to rhEPO (A) Representative flow cytometry analysis showing circulating CD71 HI /Ter119 HI reticulocytes in PHZ-treated and control mice, as well as in mice subjected to phlebotomy. For PHZ a representative experiment out of 20 is shown. For phlebotomy, a representative experiment out of 8 is shown. (B) Representative histograms of SLC3A2 expression in circulating CD71 HI /Ter119 HI reticulocytes of untreated control mice (in blue), mice treated with PHZ or mice subjected to phlebotomy (in red), as well as ErGFPcre Slc7a5 LoxP/LoxP mice treated with PHZ or subjected to phlebotomy (in green). The IgG-APC control is represented in gray. Quantification of the mean fluorescence intensity (MFI) of SLC3A2 on circulating CD71 HI /Ter119 HI reticulocytes from control untreated mice (n = 33), control mice treated with PHZ (n = 20), ErGFPcre Slc7a5 LoxP/+ untreated mice (n = 11), ErGFPcre Slc7a5 LoxP/+ mice treated with PHZ (n = 10), ErGFPcre Slc7a5 LoxP/LoxP untreated mice (n = 16) and ErGFPcre Slc7a5 LoxP/LoxP mice treated with PHZ (n = 14). Quantification of the mean fluorescence intensity (MFI) of SLC3A2 on circulating CD71 HI /Ter119 HI reticulocytes from control mice (n = 8), control mice subjected to phlebotomy (n = 8), ErGFPcre Slc7a5 LoxP/+ untreated mice (n = 4), ErGFPcre Slc7a5 LoxP/+ mice subjected to phlebotomy (n = 4), ErGFPcre Slc7a5 LoxP / LoxP untreated mice (n = 3) and ErGFPcre Slc7a5 LoxP/LoxP mice subjected to phlebotomy (n = 3). (C) Western blot analyses of SLC7A5 relative to β-actin protein levels in circulating erythroid cells of control and ErGFPcre Slc7a5 LoxP/LoxP PHZ-treated mice. (D) L-phenylalanine uptake in isolated CD71 HI /Ter119 HI circulating reticulocytes from PHZ-treated control or ErGFPcre Slc7a5 LoxP/LoxP mice (n = 5). (E) Representative flow cytometry analysis of CD71 expression versus GYPA in healthy and anemic subjects (left panels). Histograms of SLC3A2 expression in circulating CD71 HI /GYPA HI and CD71 NEG /GYPA HI populations of control (blue) and anemic (red) subjects (right panels). A representative experiment out of 3 anemic subjects and 3 control subjects is shown. (F) Representative histograms of SLC3A2 expression in circulating CD71 HI /Ter119 HI reticulocytes of untreated control mice (blue) or mice treated with rhEPO (200U) (red). The IgG-APC control is represented in gray. Quantification of the mean fluorescence intensity (MFI) of SLC3A2 on circulating CD71 HI /Ter119 HI reticulocytes from untreated (n = 5) or rhEPO-treated control mice (n = 5), as well as from untreated (n = 3) or rhEPO-treated ErGFPcre Slc7a5 LoxP/LoxP mice (n = 5). Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing more than two groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗∗p < 0.01; ∗∗∗p < 0.005).

Journal: iScience

Article Title: Erythroid SLC7A5/SLC3A2 amino acid carrier controls red blood cell size and maturation

doi: 10.1016/j.isci.2022.105739

Figure Lengend Snippet: SLC7A5/SLC3A2 expression in circulating CD71 HI /Ter119 HI reticulocytes under anemic conditions and in response to rhEPO (A) Representative flow cytometry analysis showing circulating CD71 HI /Ter119 HI reticulocytes in PHZ-treated and control mice, as well as in mice subjected to phlebotomy. For PHZ a representative experiment out of 20 is shown. For phlebotomy, a representative experiment out of 8 is shown. (B) Representative histograms of SLC3A2 expression in circulating CD71 HI /Ter119 HI reticulocytes of untreated control mice (in blue), mice treated with PHZ or mice subjected to phlebotomy (in red), as well as ErGFPcre Slc7a5 LoxP/LoxP mice treated with PHZ or subjected to phlebotomy (in green). The IgG-APC control is represented in gray. Quantification of the mean fluorescence intensity (MFI) of SLC3A2 on circulating CD71 HI /Ter119 HI reticulocytes from control untreated mice (n = 33), control mice treated with PHZ (n = 20), ErGFPcre Slc7a5 LoxP/+ untreated mice (n = 11), ErGFPcre Slc7a5 LoxP/+ mice treated with PHZ (n = 10), ErGFPcre Slc7a5 LoxP/LoxP untreated mice (n = 16) and ErGFPcre Slc7a5 LoxP/LoxP mice treated with PHZ (n = 14). Quantification of the mean fluorescence intensity (MFI) of SLC3A2 on circulating CD71 HI /Ter119 HI reticulocytes from control mice (n = 8), control mice subjected to phlebotomy (n = 8), ErGFPcre Slc7a5 LoxP/+ untreated mice (n = 4), ErGFPcre Slc7a5 LoxP/+ mice subjected to phlebotomy (n = 4), ErGFPcre Slc7a5 LoxP / LoxP untreated mice (n = 3) and ErGFPcre Slc7a5 LoxP/LoxP mice subjected to phlebotomy (n = 3). (C) Western blot analyses of SLC7A5 relative to β-actin protein levels in circulating erythroid cells of control and ErGFPcre Slc7a5 LoxP/LoxP PHZ-treated mice. (D) L-phenylalanine uptake in isolated CD71 HI /Ter119 HI circulating reticulocytes from PHZ-treated control or ErGFPcre Slc7a5 LoxP/LoxP mice (n = 5). (E) Representative flow cytometry analysis of CD71 expression versus GYPA in healthy and anemic subjects (left panels). Histograms of SLC3A2 expression in circulating CD71 HI /GYPA HI and CD71 NEG /GYPA HI populations of control (blue) and anemic (red) subjects (right panels). A representative experiment out of 3 anemic subjects and 3 control subjects is shown. (F) Representative histograms of SLC3A2 expression in circulating CD71 HI /Ter119 HI reticulocytes of untreated control mice (blue) or mice treated with rhEPO (200U) (red). The IgG-APC control is represented in gray. Quantification of the mean fluorescence intensity (MFI) of SLC3A2 on circulating CD71 HI /Ter119 HI reticulocytes from untreated (n = 5) or rhEPO-treated control mice (n = 5), as well as from untreated (n = 3) or rhEPO-treated ErGFPcre Slc7a5 LoxP/LoxP mice (n = 5). Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing more than two groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗∗p < 0.01; ∗∗∗p < 0.005).

Article Snippet: The membranes were then blocked and probed with antibodies against: LAT1 (SLC7A5) (ANT-105, Alomone Labs, Jerusalem, Israel); β-actin (A3854, Sigma-Aldrich).

Techniques: Expressing, Flow Cytometry, Control, Fluorescence, Western Blot, Isolation, Two Tailed Test

Mice lacking SLC7A5 in the erythroid lineage develop hypererythropoietinemia and smaller circulating RBCs (A) Serum EPO protein (pg/mL) in control mice (n = 18), ErGFPcre Slc7a5 LoxP/+ mice (n = 4), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 13) under baseline conditions; in control mice (n = 18), ErGFPcre Slc7a5 LoxP/+ mice (n = 9) and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 16) after PHZ treatment; and in phlebotomized control mice (n = 6), ErGFPcre Slc7a5 LoxP/+ mice (n = 4) and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 6). (B) Total number of circulating RBCs in control mice (n = 28), ErGFPcre Slc7a5 LoxP/+ mice (n = 13), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 18) under basal conditions or in control mice (n = 13), ErGFPcre Slc7a5 LoxP/+ mice (n = 9), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 13) after PHZ treatment. (C) Mean corpuscular volume (MCV, μm 3 ) of circulating RBCs in control mice (n = 28), ErGFPcre Slc7a5 LoxP/+ mice (n = 13), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 18) under basal conditions, and in control mice (n = 11), ErGFPcre Slc7a5 LoxP/+ mice (n = 7), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 12) after PHZ treatment. (D) Representative images of life imaging morphology cytometry (left) and 3D-cell rendering images (right) of RBCs of ErGFPcre Slc7a5 LoxP/LoxP and control mice. The RBC populations normally distributed their areas around a central average smaller for the ErGFPcre Slc7a5 LoxP / LoxP specimens than for control mice (central panels). Graph panels show the diameter (μm), area (μm 2 ), and elongation (%) of RBCs in control, and ErGFPcre Slc7a5 LoxP/LoxP mice. Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing three groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗p < 0.05; ∗∗∗p < 0.005).

Journal: iScience

Article Title: Erythroid SLC7A5/SLC3A2 amino acid carrier controls red blood cell size and maturation

doi: 10.1016/j.isci.2022.105739

Figure Lengend Snippet: Mice lacking SLC7A5 in the erythroid lineage develop hypererythropoietinemia and smaller circulating RBCs (A) Serum EPO protein (pg/mL) in control mice (n = 18), ErGFPcre Slc7a5 LoxP/+ mice (n = 4), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 13) under baseline conditions; in control mice (n = 18), ErGFPcre Slc7a5 LoxP/+ mice (n = 9) and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 16) after PHZ treatment; and in phlebotomized control mice (n = 6), ErGFPcre Slc7a5 LoxP/+ mice (n = 4) and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 6). (B) Total number of circulating RBCs in control mice (n = 28), ErGFPcre Slc7a5 LoxP/+ mice (n = 13), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 18) under basal conditions or in control mice (n = 13), ErGFPcre Slc7a5 LoxP/+ mice (n = 9), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 13) after PHZ treatment. (C) Mean corpuscular volume (MCV, μm 3 ) of circulating RBCs in control mice (n = 28), ErGFPcre Slc7a5 LoxP/+ mice (n = 13), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 18) under basal conditions, and in control mice (n = 11), ErGFPcre Slc7a5 LoxP/+ mice (n = 7), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 12) after PHZ treatment. (D) Representative images of life imaging morphology cytometry (left) and 3D-cell rendering images (right) of RBCs of ErGFPcre Slc7a5 LoxP/LoxP and control mice. The RBC populations normally distributed their areas around a central average smaller for the ErGFPcre Slc7a5 LoxP / LoxP specimens than for control mice (central panels). Graph panels show the diameter (μm), area (μm 2 ), and elongation (%) of RBCs in control, and ErGFPcre Slc7a5 LoxP/LoxP mice. Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing three groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗p < 0.05; ∗∗∗p < 0.005).

Article Snippet: The membranes were then blocked and probed with antibodies against: LAT1 (SLC7A5) (ANT-105, Alomone Labs, Jerusalem, Israel); β-actin (A3854, Sigma-Aldrich).

Techniques: Control, Imaging, Cytometry, Two Tailed Test

Erythroid-SLC7A5 deficient mice have red blood cells with lower hemoglobin content (A) Mean corpuscular hemoglobin (MCH, pg) of circulating RBCs in control mice (n = 28), ErGFPcre Slc7a5 LoxP/+ mice (n = 13), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 18) under basal conditions. (B) Representative peripheral blood stainings with May-Grünwald-Giemsa of ErGFPcre Slc7a5 LoxP/LoxP (right) and control (left) mice. (C) Representative flow cytometry histograms comparing the phospho-rpS6 Ser235/6 signal in CD71 HI /Ter119 HI splenic erythroblasts of control and temsirolimus-treated mice (left panel). Representative flow cytometry histogram comparing the phospho-rpS6 Ser235/6 signal in CD71 HI /Ter119 HI splenic erythroblasts in control, and ErGFPcre Slc7a5 LoxP/LoxP mice (right panel). Quantification of phospho-rpS6 Ser235/6 signal from control (n = 6) and temsirolimus-treated mice (n = 5) (left panel). Quantification of phospho-rpS6 Ser235/6 signal from control (n = 9), ErGFPcre Slc7a5 LoxP/+ mice (n = 8), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 7) (right panel). The dashed line represents unlabeled permeabilized cells. Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing three groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗p < 0.05; ∗∗∗p < 0.005).

Journal: iScience

Article Title: Erythroid SLC7A5/SLC3A2 amino acid carrier controls red blood cell size and maturation

doi: 10.1016/j.isci.2022.105739

Figure Lengend Snippet: Erythroid-SLC7A5 deficient mice have red blood cells with lower hemoglobin content (A) Mean corpuscular hemoglobin (MCH, pg) of circulating RBCs in control mice (n = 28), ErGFPcre Slc7a5 LoxP/+ mice (n = 13), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 18) under basal conditions. (B) Representative peripheral blood stainings with May-Grünwald-Giemsa of ErGFPcre Slc7a5 LoxP/LoxP (right) and control (left) mice. (C) Representative flow cytometry histograms comparing the phospho-rpS6 Ser235/6 signal in CD71 HI /Ter119 HI splenic erythroblasts of control and temsirolimus-treated mice (left panel). Representative flow cytometry histogram comparing the phospho-rpS6 Ser235/6 signal in CD71 HI /Ter119 HI splenic erythroblasts in control, and ErGFPcre Slc7a5 LoxP/LoxP mice (right panel). Quantification of phospho-rpS6 Ser235/6 signal from control (n = 6) and temsirolimus-treated mice (n = 5) (left panel). Quantification of phospho-rpS6 Ser235/6 signal from control (n = 9), ErGFPcre Slc7a5 LoxP/+ mice (n = 8), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 7) (right panel). The dashed line represents unlabeled permeabilized cells. Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing three groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗p < 0.05; ∗∗∗p < 0.005).

Article Snippet: The membranes were then blocked and probed with antibodies against: LAT1 (SLC7A5) (ANT-105, Alomone Labs, Jerusalem, Israel); β-actin (A3854, Sigma-Aldrich).

Techniques: Control, Flow Cytometry, Two Tailed Test

Loss of SLC7A5 in the erythroid lineage increases the transition between circulating reticulocytes and mature erythrocytes (A) Flow cytometry histogram comparing the CD71 expressed by circulating Ter119 positive cells in a control, and ErGFPcre Slc7a5 LoxP/LoxP mice after PHZ treatment. Quantification of the proportion of CD71 HI /Ter119 HI , CD71 INT /Ter119 HI , and CD71 NEG /Ter119 HI in control (n = 35), ErGFPcre Slc7a5 LoxP/+ mice (n = 19), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 28) after PHZ treatment. The range of CD71 expression considered for the quantification of CD71 HI /Ter119 HI , CD71 INT /Ter119 HI , and CD71 NEG /Ter119 HI RBCs is indicated. (B) Flow cytometry histogram comparing the TMRM signal in circulating CD71 HI /Ter119 HI reticulocytes of control, and ErGFPcre Slc7a5 LoxP/LoxP mice after PHZ treatment. Quantification of TMRM signal from control (n = 7), ErGFPcre Slc7a5 LoxP/+ mice (n = 6), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 7) after PHZ treatment. The dashed line represents unlabeled cells. (C) Maximal mitochondrial oxygen consumption rate (OCR) in circulating cells of PHZ-treated mice (n = 14) and PHZ-treated ErGFPcre Slc7a5 LoxP/LoxP mice (n = 15). Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing three groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.005).

Journal: iScience

Article Title: Erythroid SLC7A5/SLC3A2 amino acid carrier controls red blood cell size and maturation

doi: 10.1016/j.isci.2022.105739

Figure Lengend Snippet: Loss of SLC7A5 in the erythroid lineage increases the transition between circulating reticulocytes and mature erythrocytes (A) Flow cytometry histogram comparing the CD71 expressed by circulating Ter119 positive cells in a control, and ErGFPcre Slc7a5 LoxP/LoxP mice after PHZ treatment. Quantification of the proportion of CD71 HI /Ter119 HI , CD71 INT /Ter119 HI , and CD71 NEG /Ter119 HI in control (n = 35), ErGFPcre Slc7a5 LoxP/+ mice (n = 19), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 28) after PHZ treatment. The range of CD71 expression considered for the quantification of CD71 HI /Ter119 HI , CD71 INT /Ter119 HI , and CD71 NEG /Ter119 HI RBCs is indicated. (B) Flow cytometry histogram comparing the TMRM signal in circulating CD71 HI /Ter119 HI reticulocytes of control, and ErGFPcre Slc7a5 LoxP/LoxP mice after PHZ treatment. Quantification of TMRM signal from control (n = 7), ErGFPcre Slc7a5 LoxP/+ mice (n = 6), and ErGFPcre Slc7a5 LoxP/LoxP mice (n = 7) after PHZ treatment. The dashed line represents unlabeled cells. (C) Maximal mitochondrial oxygen consumption rate (OCR) in circulating cells of PHZ-treated mice (n = 14) and PHZ-treated ErGFPcre Slc7a5 LoxP/LoxP mice (n = 15). Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test when comparing three groups or two-tailed Student’s t test with Welch’s correction when comparing two groups (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.005).

Article Snippet: The membranes were then blocked and probed with antibodies against: LAT1 (SLC7A5) (ANT-105, Alomone Labs, Jerusalem, Israel); β-actin (A3854, Sigma-Aldrich).

Techniques: Flow Cytometry, Control, Expressing, Two Tailed Test

Journal: iScience

Article Title: Erythroid SLC7A5/SLC3A2 amino acid carrier controls red blood cell size and maturation

doi: 10.1016/j.isci.2022.105739

Figure Lengend Snippet:

Article Snippet: The membranes were then blocked and probed with antibodies against: LAT1 (SLC7A5) (ANT-105, Alomone Labs, Jerusalem, Israel); β-actin (A3854, Sigma-Aldrich).

Techniques: Control, Recombinant, Enzyme-linked Immunosorbent Assay, Knock-In, Amplification, Software

( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of Slc7a5 mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A ) Schematic diagram of the procedure for [ 125 I]IMT uptake assay. ( B and C ) Brain tissues from wild-type mice at 8–12 weeks of age were incubated with [ 125 I]IMT at 4°C or 37°C for 30 minutes in Hanks’ balanced salt solution (HBSS) buffer ( B ) ( n = 5, *** P < 0.001, 2-tailed Student’s t test) or at 37°C for 30 minutes in HBSS buffer containing 30 μM JPH203 ( C ) ( n = 10 or 11, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Quantification of mRNAs encoding LAT family in VMH ( D ) and ARC ( E ) of mice fed a normal chow diet (NC) at 8–12 weeks of age ( n = 4, * P < 0.05, ** P < 0.01, Kruskal-Wallis test post hoc Dunn’s test). TPM, transcripts per million. ( F ) Quantification of mRNAs encoding LAT family in human hypothalamus ( n = 202, *** P < 0.001, Kruskal-Wallis test post hoc Dunn’s test). ( G ) Brain tissues from WT mice fed an NC or HFD (for 17 weeks, from 7 to 24 weeks of age) at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, * P < 0.05, 2-tailed Student’s t test). ( H ) Brain tissues from WT mice and db/db mice at 24 weeks of age were incubated with [ 125 I]IMT at 37°C for 30 minutes in HBSS buffer ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( I ) LAT1-dependent [ 125 I]IMT uptake in brain tissues from WT mice and db/db mice at 24 weeks of age ( n = 5, ** P < 0.01, 2-tailed Student’s t test). ( J and K ) Quantification of Slc7a5 mRNA in VMH ( J ) and ARC ( K ) of mice fed an NC or HFD (from 5–6 to 8–12 weeks of age) at 8–12 weeks of age ( n = 4, * P < 0.05, *** P < 0.001, Fisher’s exact test). All the mice used in this study were male. CPM, counts per million.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Incubation

( A ) Weekly body weight is shown for LepR-Cre Slc7a5 fl/fl mice and control mice fed an NC ( n = 8 or 9, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA with Bonferroni post hoc test). ( B ) Gross appearance at 24 weeks of age is shown for LepR-Cre Slc7a5 fl/fl mice and control mice fed an NC. Scale bar, 1 cm. ( C – E ) Representative pictures of visceral and subcutaneous WAT (vWAT and sWAT) ( C ), adipose tissue weights ( D ), and adipose tissue weights normalized to body weight ( E ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 17 or 18, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). Scale bar, 1 cm. ( F and G ) H&E stain was performed on the vWAT of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age, followed by quantification of adipocyte size of vWAT ( n = 15 or 16, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA with Bonferroni post hoc test). Scale bar, 100 μm. ( H ) Serum leptin levels were measured in LepR-Cre Slc7a5 fl/fl mice at 22–24 weeks of age ( n = 4, *** P < 0.001, 2-tailed Student’s t test). ( I ) [ 125 I]IMT uptake in hypothalamus from LepR-Cre Slc7a5 fl/fl mice at 22–24 weeks of age ( n = 6 or 7, * P < 0.05, 2-tailed Student’s t test). ( J ) Log 2 ratio of the amino acid levels in VMH between LepR-Cre Slc7a5 fl/fl mice and control mice at 7 weeks of age ( n = 12, the amino acids with P < 0.05 are represented in red, 2-tailed Student’s t test). ( K ) Quantification of Slc7a5 mRNA in ARC and VMH of mice fed an NC ( n = 4, ** P < 0.01, Fisher’s exact test). ( L and M ) The enrichment plots for amino acid transport-related gene sets in VMH and ARC of mice fed an NC ( n = 4). All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A ) Weekly body weight is shown for LepR-Cre Slc7a5 fl/fl mice and control mice fed an NC ( n = 8 or 9, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA with Bonferroni post hoc test). ( B ) Gross appearance at 24 weeks of age is shown for LepR-Cre Slc7a5 fl/fl mice and control mice fed an NC. Scale bar, 1 cm. ( C – E ) Representative pictures of visceral and subcutaneous WAT (vWAT and sWAT) ( C ), adipose tissue weights ( D ), and adipose tissue weights normalized to body weight ( E ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 17 or 18, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). Scale bar, 1 cm. ( F and G ) H&E stain was performed on the vWAT of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age, followed by quantification of adipocyte size of vWAT ( n = 15 or 16, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA with Bonferroni post hoc test). Scale bar, 100 μm. ( H ) Serum leptin levels were measured in LepR-Cre Slc7a5 fl/fl mice at 22–24 weeks of age ( n = 4, *** P < 0.001, 2-tailed Student’s t test). ( I ) [ 125 I]IMT uptake in hypothalamus from LepR-Cre Slc7a5 fl/fl mice at 22–24 weeks of age ( n = 6 or 7, * P < 0.05, 2-tailed Student’s t test). ( J ) Log 2 ratio of the amino acid levels in VMH between LepR-Cre Slc7a5 fl/fl mice and control mice at 7 weeks of age ( n = 12, the amino acids with P < 0.05 are represented in red, 2-tailed Student’s t test). ( K ) Quantification of Slc7a5 mRNA in ARC and VMH of mice fed an NC ( n = 4, ** P < 0.01, Fisher’s exact test). ( L and M ) The enrichment plots for amino acid transport-related gene sets in VMH and ARC of mice fed an NC ( n = 4). All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Control, Staining

( A ) Glucose tolerance tests (GTTs) were performed in LepR-Cre Slc7a5 fl/fl mice and control mice after a 6-hour fast at 22–24 weeks of age ( n = 6 to 8, 2-way ANOVA with Bonferroni post hoc test). ( B ) Insulin tolerance tests (ITTs) were performed in LepR-Cre Slc7a5 fl/fl mice and control mice after a 6-hour fast at 22–24 weeks of age ( n = 10 or 11, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA with Bonferroni post hoc test). ( C ) Serum insulin levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice after a 6-hour fast at 22–24 weeks of age ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Immunohistochemical stain for insulin was performed on the islets of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age, followed by quantification of insulin-positive area ( n = 6 to 10, * P < 0.05, 2-tailed Student’s t test). Scale bar, 200 μm. ( F – H ) Representative picture of liver ( F ), tissue weights ( G ), and tissue weights normalized to body weight ( H ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 17 or 18, *** P < 0.001, 2-tailed Student’s t test). Scale bar, 500 μm. ( I ) Oil Red O stain was performed on the liver of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age. Scale bar, 100 μm. All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A ) Glucose tolerance tests (GTTs) were performed in LepR-Cre Slc7a5 fl/fl mice and control mice after a 6-hour fast at 22–24 weeks of age ( n = 6 to 8, 2-way ANOVA with Bonferroni post hoc test). ( B ) Insulin tolerance tests (ITTs) were performed in LepR-Cre Slc7a5 fl/fl mice and control mice after a 6-hour fast at 22–24 weeks of age ( n = 10 or 11, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-way ANOVA with Bonferroni post hoc test). ( C ) Serum insulin levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice after a 6-hour fast at 22–24 weeks of age ( n = 5, *** P < 0.001, 2-tailed Student’s t test). ( D and E ) Immunohistochemical stain for insulin was performed on the islets of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age, followed by quantification of insulin-positive area ( n = 6 to 10, * P < 0.05, 2-tailed Student’s t test). Scale bar, 200 μm. ( F – H ) Representative picture of liver ( F ), tissue weights ( G ), and tissue weights normalized to body weight ( H ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 17 or 18, *** P < 0.001, 2-tailed Student’s t test). Scale bar, 500 μm. ( I ) Oil Red O stain was performed on the liver of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age. Scale bar, 100 μm. All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Control, Immunohistochemical staining, Staining

( A – F ) Food intake ( A ), locomotor activity ( B ), O 2 consumption ( C ), CO 2 production ( D ), energy expenditure ( E ), and body temperature ( F ) were measured in singly housed LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 7 to 10, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). ( G – I ) Representative picture of BAT ( G ), tissue weights ( H ), and tissue weights normalized to body weight ( I ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 17 or 18, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). Scale bar, 1 cm. ( J ) H&E stain was performed on the BAT of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age. Scale bar, 100 μm. ( K ) Transmission electron microscopy analysis was performed on the BAT of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age. Scale bar, 10 μm. ( L and M ) mtDNA content of BAT ( L ) and mRNA expression ( M ) were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 3 to 4, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A – F ) Food intake ( A ), locomotor activity ( B ), O 2 consumption ( C ), CO 2 production ( D ), energy expenditure ( E ), and body temperature ( F ) were measured in singly housed LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 7 to 10, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). ( G – I ) Representative picture of BAT ( G ), tissue weights ( H ), and tissue weights normalized to body weight ( I ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 17 or 18, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). Scale bar, 1 cm. ( J ) H&E stain was performed on the BAT of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age. Scale bar, 100 μm. ( K ) Transmission electron microscopy analysis was performed on the BAT of LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age. Scale bar, 10 μm. ( L and M ) mtDNA content of BAT ( L ) and mRNA expression ( M ) were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 3 to 4, * P < 0.05, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Activity Assay, Control, Staining, Transmission Assay, Electron Microscopy, Expressing

( A and B ) Serum epinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( A ) or 24 ( B ) weeks of age (7 wk; n = 8, 24 wk; n = 4, ** P < 0.01, 2-tailed Student’s t test). ( C and D ) Serum norepinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( C ) or 24 ( D ) weeks of age (7 wk; n = 8, 24 wk; n = 4, *** P < 0.001, 2-tailed Student’s t test). ( E and F ) Urine epinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( E ) or 24 ( F ) weeks of age (7 wk; n = 8, 24 wk; n = 4, ** P < 0.01, 2-tailed Student’s t test). ( G and H ) Urine norepinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( G ) or 24 ( H ) weeks of age (7 wk; n = 8, 24 wk; n = 4, * P < 0.05, 2-tailed Student’s t test). ( I and J ) Norepinephrine turnover levels of ( I ) BAT and ( J ) soleus muscle were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 24 weeks of age ( n = 8 to 10, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). ( K ) Serum leptin levels were measured in LepR-Cre Slc7a5 fl/fl mice at 7 weeks of age ( n = 8, ** P < 0.01, 2-tailed Student’s t test). ( L – O ) Immunohistochemical analysis of phosphorylated STAT3 activation 2 hours after intraperitoneal injection of 5 mg/kg leptin was performed on the VMH ( L and M ) and ARC ( N and O ) of LepR-Cre Slc7a5 fl/fl mice and control mice at 7 weeks of age ( n = 3 or 4, * P < 0.05, ** P < 0.01, # P < 0.05, 2-way ANOVA with Bonferroni post hoc test). Scale bar, 100 μm. Arrowheads indicate representative phosphorylated STAT3/tdTomato double-positive neurons. All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A and B ) Serum epinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( A ) or 24 ( B ) weeks of age (7 wk; n = 8, 24 wk; n = 4, ** P < 0.01, 2-tailed Student’s t test). ( C and D ) Serum norepinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( C ) or 24 ( D ) weeks of age (7 wk; n = 8, 24 wk; n = 4, *** P < 0.001, 2-tailed Student’s t test). ( E and F ) Urine epinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( E ) or 24 ( F ) weeks of age (7 wk; n = 8, 24 wk; n = 4, ** P < 0.01, 2-tailed Student’s t test). ( G and H ) Urine norepinephrine levels were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 7 ( G ) or 24 ( H ) weeks of age (7 wk; n = 8, 24 wk; n = 4, * P < 0.05, 2-tailed Student’s t test). ( I and J ) Norepinephrine turnover levels of ( I ) BAT and ( J ) soleus muscle were measured in LepR-Cre Slc7a5 fl/fl mice and control mice at 24 weeks of age ( n = 8 to 10, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test). ( K ) Serum leptin levels were measured in LepR-Cre Slc7a5 fl/fl mice at 7 weeks of age ( n = 8, ** P < 0.01, 2-tailed Student’s t test). ( L – O ) Immunohistochemical analysis of phosphorylated STAT3 activation 2 hours after intraperitoneal injection of 5 mg/kg leptin was performed on the VMH ( L and M ) and ARC ( N and O ) of LepR-Cre Slc7a5 fl/fl mice and control mice at 7 weeks of age ( n = 3 or 4, * P < 0.05, ** P < 0.01, # P < 0.05, 2-way ANOVA with Bonferroni post hoc test). Scale bar, 100 μm. Arrowheads indicate representative phosphorylated STAT3/tdTomato double-positive neurons. All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Control, Immunohistochemical staining, Activation Assay, Injection

( A ) Micro–computed tomography (μCT) analysis (scale bar, 1 mm); ( B ) bone volume over tissue volume (BV/TV) ratio as determined by μCT ( n = 12 or 13, * P < 0.05, 2-tailed Student’s t test); ( C ) von Kossa stain, scale bar, 1 mm; ( D ) BV/TV ratio as determined by von Kossa stain ( n = 5 or 6, * P < 0.05, 2-tailed Student’s t test); ( E ) number of osteoblasts/tissue area ratio ( n = 5 or 6, ** P < 0.01, 2-tailed Student’s t test); ( F ) calcein labeling (scale bar, 50 μm); ( G ) bone formation rate ( n = 5 or 6, * P < 0.05, 2-tailed Student’s t test); ( H ) TRAP stain (scale bar, 50 μm; arrowheads indicate TRAP-positive osteoclasts); and ( I ) osteoclast surface/bone surface ratio of femurs from LepR-Cre Slc7a5 fl/fl mice and control mice at 12–16 weeks of age ( n = 3 to 5, ** P < 0.01, 2-tailed Student’s t test). TRAP, tartrate-resistant acid phosphatase. ( J ) μCT analysis, scale bar, 1 mm and ( K ) BV/TV ratio as determined by μCT of femurs from LepR-Cre Slc7a5 fl/fl mice administrated with isoproterenol at 14 weeks of age ( n = 10, ** P < 0.01, # P < 0.05, 2-tailed Student’s t test with Benjamini-Hochberg correction). ( L ) Representative images of CFU assays stained with crystal violet and alizarin red (scale bar, 500 μm), ( M ) mRNA level of Slc7a5 in BM-MSCs ( n = 4, ** P < 0.01, 2-tailed Student’s t test), and ( N and O ) quantification of CFU assays stained with crystal violet ( N ) and alizarin red ( O ) ( n = 6 to 9, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test) in LepR-Cre Slc7a5 fl/fl mice at 6 weeks of age. All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A ) Micro–computed tomography (μCT) analysis (scale bar, 1 mm); ( B ) bone volume over tissue volume (BV/TV) ratio as determined by μCT ( n = 12 or 13, * P < 0.05, 2-tailed Student’s t test); ( C ) von Kossa stain, scale bar, 1 mm; ( D ) BV/TV ratio as determined by von Kossa stain ( n = 5 or 6, * P < 0.05, 2-tailed Student’s t test); ( E ) number of osteoblasts/tissue area ratio ( n = 5 or 6, ** P < 0.01, 2-tailed Student’s t test); ( F ) calcein labeling (scale bar, 50 μm); ( G ) bone formation rate ( n = 5 or 6, * P < 0.05, 2-tailed Student’s t test); ( H ) TRAP stain (scale bar, 50 μm; arrowheads indicate TRAP-positive osteoclasts); and ( I ) osteoclast surface/bone surface ratio of femurs from LepR-Cre Slc7a5 fl/fl mice and control mice at 12–16 weeks of age ( n = 3 to 5, ** P < 0.01, 2-tailed Student’s t test). TRAP, tartrate-resistant acid phosphatase. ( J ) μCT analysis, scale bar, 1 mm and ( K ) BV/TV ratio as determined by μCT of femurs from LepR-Cre Slc7a5 fl/fl mice administrated with isoproterenol at 14 weeks of age ( n = 10, ** P < 0.01, # P < 0.05, 2-tailed Student’s t test with Benjamini-Hochberg correction). ( L ) Representative images of CFU assays stained with crystal violet and alizarin red (scale bar, 500 μm), ( M ) mRNA level of Slc7a5 in BM-MSCs ( n = 4, ** P < 0.01, 2-tailed Student’s t test), and ( N and O ) quantification of CFU assays stained with crystal violet ( N ) and alizarin red ( O ) ( n = 6 to 9, ** P < 0.01, *** P < 0.001, 2-tailed Student’s t test) in LepR-Cre Slc7a5 fl/fl mice at 6 weeks of age. All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Micro-CT, Staining, Labeling, Control

( A ) Schematic diagram of the bilateral viral microinjection into the VMH and representative validation image of mCherry expression in the VMH. Scale bar, 500 μm. ( B ) Weekly body weight after injection of AAV- Control or AAV- Slc7a5 into the VMH is shown for LepR-Cre Slc7a5 fl/fl mice and control mice ( n = 8 to 12, ** P < 0.01, *** P < 0.001: versus LepR-Cre /AAV- Control , # P < 0.05, ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl /AAV- Control , 2-way ANOVA with Bonferroni post hoc test). ( C and D ) Adipose tissue weights ( C ) and adipose tissue weights normalized to body weight ( D ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice injected with AAV- Control or AAV- Slc7a5 into the VMH at 16 weeks of age ( n = 8, ** P < 0.01, *** P < 0.001, # P < 0.05, ## P < 0.01, ### P < 0.001, 2-tailed Student’s t test with Bonferroni correction). ( E ) ITTs were performed in LepR-Cre Slc7a5 fl/fl mice and control mice injected with AAV- Control or AAV- Slc7a5 into the VMH after a 6-hour fast at 16 weeks of age ( n = 3 or 4, * P < 0.05, ** P < 0.01: versus LepR-Cre /AAV- Control , # P < 0.05, ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl /AAV- Control , 2-way ANOVA with Bonferroni post hoc test). ( F ) μCT analysis (scale bar, 1 mm) and ( G ) BV/TV ratio as determined by μCT of femurs from LepR-Cre Slc7a5 fl/fl mice and control mice injected with AAV- Control or AAV- Slc7a5 into the VMH at 12–16 weeks of age ( n = 6 to 11, * P < 0.05, # P < 0.05, 2-way ANOVA with Bonferroni post hoc test). All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A ) Schematic diagram of the bilateral viral microinjection into the VMH and representative validation image of mCherry expression in the VMH. Scale bar, 500 μm. ( B ) Weekly body weight after injection of AAV- Control or AAV- Slc7a5 into the VMH is shown for LepR-Cre Slc7a5 fl/fl mice and control mice ( n = 8 to 12, ** P < 0.01, *** P < 0.001: versus LepR-Cre /AAV- Control , # P < 0.05, ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl /AAV- Control , 2-way ANOVA with Bonferroni post hoc test). ( C and D ) Adipose tissue weights ( C ) and adipose tissue weights normalized to body weight ( D ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice injected with AAV- Control or AAV- Slc7a5 into the VMH at 16 weeks of age ( n = 8, ** P < 0.01, *** P < 0.001, # P < 0.05, ## P < 0.01, ### P < 0.001, 2-tailed Student’s t test with Bonferroni correction). ( E ) ITTs were performed in LepR-Cre Slc7a5 fl/fl mice and control mice injected with AAV- Control or AAV- Slc7a5 into the VMH after a 6-hour fast at 16 weeks of age ( n = 3 or 4, * P < 0.05, ** P < 0.01: versus LepR-Cre /AAV- Control , # P < 0.05, ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl /AAV- Control , 2-way ANOVA with Bonferroni post hoc test). ( F ) μCT analysis (scale bar, 1 mm) and ( G ) BV/TV ratio as determined by μCT of femurs from LepR-Cre Slc7a5 fl/fl mice and control mice injected with AAV- Control or AAV- Slc7a5 into the VMH at 12–16 weeks of age ( n = 6 to 11, * P < 0.05, # P < 0.05, 2-way ANOVA with Bonferroni post hoc test). All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Microinjection, Expressing, Injection, Control

( A and B ) Immunohistochemical analysis of pS6 activation 2 hours after intraperitoneal injection of 5 mg/kg leptin was performed on the VMH of LepR-Cre Slc7a5 fl/fl mice and control mice at 7 weeks of age ( n = 3, ** P < 0.01, # P < 0.05, 2-way ANOVA with Bonferroni post hoc test). Scale bar, 100 μm. Arrowheads indicate pS6/tdTomato double-positive neurons. ( C ) Gross appearance at 24 weeks of age is shown for LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice fed an NC. Scale bar, 1 cm. ( D ) Weekly body weight is shown for LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice fed NC ( n = 9 to 11, * P < 0.05, ** P < 0.01, *** P < 0.001: versus Slc7a5 fl/fl , # P < 0.05, ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl , 2-way ANOVA with Bonferroni post hoc test). ( E – G ) Representative pictures of vWAT, sWAT, and BAT ( E ); adipose tissue weights ( F ); and adipose tissue weights normalized to body weight ( G ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 12 to 18, ** P < 0.01, *** P < 0.001, ### P < 0.001, 1-way ANOVA with Bonferroni post hoc test). Scale bar, 1 cm. ( H ) ITTs were performed in LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice after a 6-hour fast at 22–24 weeks of age ( n = 3 to 9, * P < 0.05, ** P < 0.01: versus Slc7a5 fl/fl , ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl , 2-way ANOVA with Bonferroni post hoc test). All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A and B ) Immunohistochemical analysis of pS6 activation 2 hours after intraperitoneal injection of 5 mg/kg leptin was performed on the VMH of LepR-Cre Slc7a5 fl/fl mice and control mice at 7 weeks of age ( n = 3, ** P < 0.01, # P < 0.05, 2-way ANOVA with Bonferroni post hoc test). Scale bar, 100 μm. Arrowheads indicate pS6/tdTomato double-positive neurons. ( C ) Gross appearance at 24 weeks of age is shown for LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice fed an NC. Scale bar, 1 cm. ( D ) Weekly body weight is shown for LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice fed NC ( n = 9 to 11, * P < 0.05, ** P < 0.01, *** P < 0.001: versus Slc7a5 fl/fl , # P < 0.05, ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl , 2-way ANOVA with Bonferroni post hoc test). ( E – G ) Representative pictures of vWAT, sWAT, and BAT ( E ); adipose tissue weights ( F ); and adipose tissue weights normalized to body weight ( G ) are shown for LepR-Cre Slc7a5 fl/fl mice and control mice at 22–24 weeks of age ( n = 12 to 18, ** P < 0.01, *** P < 0.001, ### P < 0.001, 1-way ANOVA with Bonferroni post hoc test). Scale bar, 1 cm. ( H ) ITTs were performed in LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice after a 6-hour fast at 22–24 weeks of age ( n = 3 to 9, * P < 0.05, ** P < 0.01: versus Slc7a5 fl/fl , ## P < 0.01, ### P < 0.001: versus LepR-Cre Slc7a5 fl/fl , 2-way ANOVA with Bonferroni post hoc test). All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Immunohistochemical staining, Activation Assay, Injection, Control

( A ) μCT analysis, scale bar, 1 mm; and ( B ) BV/TV ratio as determined by μCT of femurs from LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice at 12–16 weeks of age ( n = 3 to 14, *** P < 0.001, # P < 0.05, 1-way ANOVA with Bonferroni post hoc test). ( C ) Working model of the findings of this study. All the mice used in this study were male.

Journal: JCI Insight

Article Title: l- Type amino acid transporter 1 in hypothalamic neurons in mice maintains energy and bone homeostasis

doi: 10.1172/jci.insight.154925

Figure Lengend Snippet: ( A ) μCT analysis, scale bar, 1 mm; and ( B ) BV/TV ratio as determined by μCT of femurs from LepR-Cre Slc7a5 fl/fl mice, LepR-Cre Slc7a5 fl/fl Tsc1 fl/+ mice, and control mice at 12–16 weeks of age ( n = 3 to 14, *** P < 0.001, # P < 0.05, 1-way ANOVA with Bonferroni post hoc test). ( C ) Working model of the findings of this study. All the mice used in this study were male.

Article Snippet: pAAV - hSynI - DIO - HA - Slc7a5 - mCherry vector was generated by replacing rM3D(Gs) cDNA in pAAV-hSyn-DIO-rM3D(Gs)-mCherry (Addgene 50458, a gift from Bryan Roth, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA) with a mouse Slc7a5 cDNA from pCMV6 - Slc7a5 (Origene, MC217270) vector. pAAV - hSynI - DIO - HA - mCherry vector was generated by deleting rM3D(Gs) cDNA from pAAV-hSyn-DIO-rM3D(Gs)-mCherry .

Techniques: Control

Schematic illustration of the work which showed biologically transformable NPs (aptamer and peptide-modified DTX-loaded pH and glutathione (GSH) dual responsive polymeric micelles) with altered performance after exposure to overexpressed MMP-9 in breast cancer TME. The synthesized NPs generally exhibit relatively neutral biological activity in the bloodstream and show minimal interaction with blood proteins (especially opsonins), as noted in the earlier article. However, upon entering the tumor microenvironment, and following the breakdown of the fusogenic peptide, the hidden aptamers (TA1) between these peptides, along with the remaining peptide sequence (SRL-2), suddenly become exposed. These aptamers alongside of SRL-2 peptide (10 amino acid remaining after cleavage of fusogenic peptide) then synergistically recognize the CD44 and LRP-1, respectively on cancer cells, demonstrating a significantly high cellular uptake efficiency. Essentially, our initial nanoparticles function like a shuttle, as depicted, that upon reaching the tumor boundary (illustrated by the gray bubble around the tumor in the schematic), loses part of its structure. The remaining part of the structure, resembling a missile cap with dual targeting ends, identifies the target cell receptors and effectively delivers the chemotherapeutic drug with high efficacy (at 1/10th of the therapeutic concentration) to the cancer cells.

Journal: Scientific Reports

Article Title: Enhanced docetaxel therapeutic effect using dual targeted SRL-2 and TA1 aptamer conjugated micelles in inhibition Balb/c mice breast cancer model

doi: 10.1038/s41598-024-75042-8

Figure Lengend Snippet: Schematic illustration of the work which showed biologically transformable NPs (aptamer and peptide-modified DTX-loaded pH and glutathione (GSH) dual responsive polymeric micelles) with altered performance after exposure to overexpressed MMP-9 in breast cancer TME. The synthesized NPs generally exhibit relatively neutral biological activity in the bloodstream and show minimal interaction with blood proteins (especially opsonins), as noted in the earlier article. However, upon entering the tumor microenvironment, and following the breakdown of the fusogenic peptide, the hidden aptamers (TA1) between these peptides, along with the remaining peptide sequence (SRL-2), suddenly become exposed. These aptamers alongside of SRL-2 peptide (10 amino acid remaining after cleavage of fusogenic peptide) then synergistically recognize the CD44 and LRP-1, respectively on cancer cells, demonstrating a significantly high cellular uptake efficiency. Essentially, our initial nanoparticles function like a shuttle, as depicted, that upon reaching the tumor boundary (illustrated by the gray bubble around the tumor in the schematic), loses part of its structure. The remaining part of the structure, resembling a missile cap with dual targeting ends, identifies the target cell receptors and effectively delivers the chemotherapeutic drug with high efficacy (at 1/10th of the therapeutic concentration) to the cancer cells.

Article Snippet: 5’- CCAAGGCCTGCAAGGGAACCAAGGACACAGTTTTTTTTTT-SH-3’ sequence which belongs to CD44 aptamer (TA1) was bought from Eurofins genomics company (Germany).

Techniques: Modification, Synthesized, Activity Assay, Sequencing, Concentration Assay