Growth characteristics of fibroblasts stably expressing AGAT and GAMT.

<p>(A) A model depicting the two sources of intracellular creatine. Creatine is either imported via CT1 or synthesized from its amino acid precursors: L-arginine; glycine; and methionine. The latter is converted into SAM via MAT2. Created with BioRender.com. (B) Growth of WT fibroblasts stably...

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Main Author: Chloe Wells (17231887) (author)
Other Authors: Jon Sorgenfrei (21273708) (author), Sadie L. Johnson (21273711) (author), Devin Albertson (21273714) (author), Jared Rutter (2128906) (author), Steven Andrew Baker (10150952) (author)
Published: 2025
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_version_ 1852020628567097344
author Chloe Wells (17231887)
author2 Jon Sorgenfrei (21273708)
Sadie L. Johnson (21273711)
Devin Albertson (21273714)
Jared Rutter (2128906)
Steven Andrew Baker (10150952)
author2_role author
author
author
author
author
author_facet Chloe Wells (17231887)
Jon Sorgenfrei (21273708)
Sadie L. Johnson (21273711)
Devin Albertson (21273714)
Jared Rutter (2128906)
Steven Andrew Baker (10150952)
author_role author
dc.creator.none.fl_str_mv Chloe Wells (17231887)
Jon Sorgenfrei (21273708)
Sadie L. Johnson (21273711)
Devin Albertson (21273714)
Jared Rutter (2128906)
Steven Andrew Baker (10150952)
dc.date.none.fl_str_mv 2025-05-08T17:50:49Z
dc.identifier.none.fl_str_mv 10.1371/journal.pone.0319350.g007
dc.relation.none.fl_str_mv https://figshare.com/articles/figure/Growth_characteristics_of_fibroblasts_stably_expressing_AGAT_and_GAMT_/28964394
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biochemistry
Medicine
Cell Biology
Genetics
Molecular Biology
Mental Health
target cell types
natural cellular compartment
highly energetic tissues
critical metabolite used
affected individuals suffer
creatine uptake relies
creatine transporter 1
healthy control fibroblasts
endogenous creatine synthesis
div >< p
intracellular creatine concentrations
intracellular synthesis
transport creatine
primary fibroblasts
patient fibroblasts
intracellular concentration
slc6a8 </
selectively expressed
protein product
proper targeting
phenotypes referred
linked gene
like behaviors
intellectual disability
hek293t cells
genetic defects
gene therapy
gatm </
function mutations
far exceeding
effective therapies
common set
common form
dc.title.none.fl_str_mv Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
dc.type.none.fl_str_mv Image
Figure
info:eu-repo/semantics/publishedVersion
image
description <p>(A) A model depicting the two sources of intracellular creatine. Creatine is either imported via CT1 or synthesized from its amino acid precursors: L-arginine; glycine; and methionine. The latter is converted into SAM via MAT2. Created with BioRender.com. (B) Growth of WT fibroblasts stably expressing the indicated constructs in standard media (DMEM + 15% FBS + P/S). N = 6. Two-sided K-S test n.s. indicates not significant. (C) Growth of CT1 loss-of-function (<i>SLC6A8</i><sup><i>Δex10-11/y</i></sup>) patient fibroblasts stably expressing the indicated constructs in standard media. N = 6. Two-sided K-S test **** indicates <i>P</i> < 0.0001. (D) Similar experiment to (C) except media was supplemented with creatine precursor amino acids to 1 mM above standard media. N = 6. Two-sided K-S test *** indicates <i>P</i> < 0.001. (E) Growth of CT1 loss-of-function (<i>SLC6A8</i><sup><i>W556X/y</i></sup>) patient fibroblasts stably expressing the indicated constructs in standard media. N = 6. Two-sided K-S test * indicates <i>P</i> < 0.05. (F) Similar experiment to (E) except media was supplemented with creatine precursor amino acids. N = 6. Two-sided K-S test **** indicates <i>P</i> < 0.0001. Lines indicate a LOESS curve fit to the mean at each timepoint. Error bars show mean + /- SEM.</p>
eu_rights_str_mv openAccess
id Manara_df06175cfeccd5ae81832fa78e5bf2e2
identifier_str_mv 10.1371/journal.pone.0319350.g007
network_acronym_str Manara
network_name_str ManaraRepo
oai_identifier_str oai:figshare.com:article/28964394
publishDate 2025
repository.mail.fl_str_mv
repository.name.fl_str_mv
repository_id_str
rights_invalid_str_mv CC BY 4.0
spelling Growth characteristics of fibroblasts stably expressing AGAT and GAMT.Chloe Wells (17231887)Jon Sorgenfrei (21273708)Sadie L. Johnson (21273711)Devin Albertson (21273714)Jared Rutter (2128906)Steven Andrew Baker (10150952)BiochemistryMedicineCell BiologyGeneticsMolecular BiologyMental Healthtarget cell typesnatural cellular compartmenthighly energetic tissuescritical metabolite usedaffected individuals suffercreatine uptake reliescreatine transporter 1healthy control fibroblastsendogenous creatine synthesisdiv >< pintracellular creatine concentrationsintracellular synthesistransport creatineprimary fibroblastspatient fibroblastsintracellular concentrationslc6a8 </selectively expressedprotein productproper targetingphenotypes referredlinked genelike behaviorsintellectual disabilityhek293t cellsgenetic defectsgene therapygatm </function mutationsfar exceedingeffective therapiescommon setcommon form<p>(A) A model depicting the two sources of intracellular creatine. Creatine is either imported via CT1 or synthesized from its amino acid precursors: L-arginine; glycine; and methionine. The latter is converted into SAM via MAT2. Created with BioRender.com. (B) Growth of WT fibroblasts stably expressing the indicated constructs in standard media (DMEM + 15% FBS + P/S). N = 6. Two-sided K-S test n.s. indicates not significant. (C) Growth of CT1 loss-of-function (<i>SLC6A8</i><sup><i>Δex10-11/y</i></sup>) patient fibroblasts stably expressing the indicated constructs in standard media. N = 6. Two-sided K-S test **** indicates <i>P</i> < 0.0001. (D) Similar experiment to (C) except media was supplemented with creatine precursor amino acids to 1 mM above standard media. N = 6. Two-sided K-S test *** indicates <i>P</i> < 0.001. (E) Growth of CT1 loss-of-function (<i>SLC6A8</i><sup><i>W556X/y</i></sup>) patient fibroblasts stably expressing the indicated constructs in standard media. N = 6. Two-sided K-S test * indicates <i>P</i> < 0.05. (F) Similar experiment to (E) except media was supplemented with creatine precursor amino acids. N = 6. Two-sided K-S test **** indicates <i>P</i> < 0.0001. Lines indicate a LOESS curve fit to the mean at each timepoint. Error bars show mean + /- SEM.</p>2025-05-08T17:50:49ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pone.0319350.g007https://figshare.com/articles/figure/Growth_characteristics_of_fibroblasts_stably_expressing_AGAT_and_GAMT_/28964394CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/289643942025-05-08T17:50:49Z
spellingShingle Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
Chloe Wells (17231887)
Biochemistry
Medicine
Cell Biology
Genetics
Molecular Biology
Mental Health
target cell types
natural cellular compartment
highly energetic tissues
critical metabolite used
affected individuals suffer
creatine uptake relies
creatine transporter 1
healthy control fibroblasts
endogenous creatine synthesis
div >< p
intracellular creatine concentrations
intracellular synthesis
transport creatine
primary fibroblasts
patient fibroblasts
intracellular concentration
slc6a8 </
selectively expressed
protein product
proper targeting
phenotypes referred
linked gene
like behaviors
intellectual disability
hek293t cells
genetic defects
gene therapy
gatm </
function mutations
far exceeding
effective therapies
common set
common form
status_str publishedVersion
title Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
title_full Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
title_fullStr Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
title_full_unstemmed Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
title_short Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
title_sort Growth characteristics of fibroblasts stably expressing AGAT and GAMT.
topic Biochemistry
Medicine
Cell Biology
Genetics
Molecular Biology
Mental Health
target cell types
natural cellular compartment
highly energetic tissues
critical metabolite used
affected individuals suffer
creatine uptake relies
creatine transporter 1
healthy control fibroblasts
endogenous creatine synthesis
div >< p
intracellular creatine concentrations
intracellular synthesis
transport creatine
primary fibroblasts
patient fibroblasts
intracellular concentration
slc6a8 </
selectively expressed
protein product
proper targeting
phenotypes referred
linked gene
like behaviors
intellectual disability
hek293t cells
genetic defects
gene therapy
gatm </
function mutations
far exceeding
effective therapies
common set
common form