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Cytoplasmic (Cyt.) levels of TDP-43 ( A ) and TDP-35CTF ( B ) in control (Ctrl; n ≥ 6) and serum-deprived HEK293 cells (Starvation), untreated (Untr; n ≥ 6) or treated with 1 μM HDACi (n=3-6): <t>SAHA,</t> <t>RGF109,</t> <t>RGFP966,</t> ACY-738, tubastatin A (tubA) and selisistat. Dashed line indicates Ctrl level. ( C ) PPIAK125ac levels in total lysate of control (Ctrl; n=5) and serum-deprived HEK293 cells (Starvation), untreated (Untr; n=5) or treated with 1 μM SAHA (n=6). ( D ) PPIAK125ac levels in total lysate of HEK293 transiently transfected with siRNA control (Scramble; n=3), siRNA for HDAC1 (n=4) and HDAC3 (n=4). ( A-D ) Data (mean ± SEM) indicate WB immunoreactivity normalized to total protein loading ( A-D ) and to PPIA levels ( C-D ). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA, Dunnett’s multiple comparisons ( A-B ), Uncorrected Fisher’s LSD ( C ) or Tukey’s multiple comparisons ( D ) post-hoc tests.
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Cytoplasmic (Cyt.) levels of TDP-43 ( A ) and TDP-35CTF ( B ) in control (Ctrl; n ≥ 6) and serum-deprived HEK293 cells (Starvation), untreated (Untr; n ≥ 6) or treated with 1 μM HDACi (n=3-6): SAHA, RGF109, RGFP966, ACY-738, tubastatin A (tubA) and selisistat. Dashed line indicates Ctrl level. ( C ) PPIAK125ac levels in total lysate of control (Ctrl; n=5) and serum-deprived HEK293 cells (Starvation), untreated (Untr; n=5) or treated with 1 μM SAHA (n=6). ( D ) PPIAK125ac levels in total lysate of HEK293 transiently transfected with siRNA control (Scramble; n=3), siRNA for HDAC1 (n=4) and HDAC3 (n=4). ( A-D ) Data (mean ± SEM) indicate WB immunoreactivity normalized to total protein loading ( A-D ) and to PPIA levels ( C-D ). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA, Dunnett’s multiple comparisons ( A-B ), Uncorrected Fisher’s LSD ( C ) or Tukey’s multiple comparisons ( D ) post-hoc tests.

Journal: bioRxiv

Article Title: Lysine deacetylation inhibition reverses TDP-43 mislocalization and in combination with arimoclomol ameliorates neuromuscular pathology

doi: 10.1101/2024.12.15.628528

Figure Lengend Snippet: Cytoplasmic (Cyt.) levels of TDP-43 ( A ) and TDP-35CTF ( B ) in control (Ctrl; n ≥ 6) and serum-deprived HEK293 cells (Starvation), untreated (Untr; n ≥ 6) or treated with 1 μM HDACi (n=3-6): SAHA, RGF109, RGFP966, ACY-738, tubastatin A (tubA) and selisistat. Dashed line indicates Ctrl level. ( C ) PPIAK125ac levels in total lysate of control (Ctrl; n=5) and serum-deprived HEK293 cells (Starvation), untreated (Untr; n=5) or treated with 1 μM SAHA (n=6). ( D ) PPIAK125ac levels in total lysate of HEK293 transiently transfected with siRNA control (Scramble; n=3), siRNA for HDAC1 (n=4) and HDAC3 (n=4). ( A-D ) Data (mean ± SEM) indicate WB immunoreactivity normalized to total protein loading ( A-D ) and to PPIA levels ( C-D ). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA, Dunnett’s multiple comparisons ( A-B ), Uncorrected Fisher’s LSD ( C ) or Tukey’s multiple comparisons ( D ) post-hoc tests.

Article Snippet: Drugs for cells and mice treatments were purchased from MedChemExpress and were as follows: SAHA (HY-10221), RGFP109 (HY-16425), RGFP966 (HY-13909); ACY-738 (HY-19327); tubastatin A (HY-13271A), selisistat (HY-15452), arimoclomol maleate (HY-106443A).

Techniques: Control, Transfection