Journal: bioRxiv

Article Title: Decoupled calcium homeostasis and signaling associated with cytoskeletal instability in YWHAG R132C induced pluripotent stem cell-derived cortical neurons

doi: 10.64898/2026.04.01.715876

Figure Lengend Snippet: a) YWHAG R132C/+ neurons have an elevated calcium baseline when compared to isogenic controls ( YWHAG +/+ normalized z-score Mean ± S.E.M = −0.63 ± 0.124, YWHAG R132C/+ = 0.713 ± 0.0730; LME accounting for batch, p<2×10 −16 , n=3 independent biological replicates). Exposure to lovastatin (10 μM) for 1h and 30m was sufficient to lower the calcium baseline, both in YWHAG R132C/+ and control iPSC lines ( YWHAG +/+ normalized z-score Mean ± S.E.M = −1.05 ± 0.154, YWHAG R132C/+ = 0.286 ± 0.162; LME accounting for batch, p=0.00412, n=3 independent biological replicates, no significant genotype-treatment interaction). b) Frequency (Hz) of calcium spikes was not altered by genotype ( YWHAG +/+ Mean frequency ± S.E.M = 3.45 ± 0.485; YWHAG R132C/+ = 3.02 ± 0.248; LME accounting for batch, p=0.09, n=3 independent biological replicates) but lovastatin treatment reduced the frequency in both genotypes ( YWHAG +/+ Mean frequency ± S.E.M = 2.36 ± 0.210; YWHAG R132C/+ = 2.45 ± 0.264; LME accounting for batch, p=1.2×10 −4 ; n=3 independent biological replicates); and c, d) there was a reduction in the amplitude of calcium spikes in YWHAG R132C/+ ( YWHAG +/+ Mean amplitude ± S.E.M = 0.0239 ± 0.00170; YWHAG R132C/+ = 0.0201 ± 0.000343; LME accounting for batch, p=3.1×10 −8 ; n=3 independent biological replicates), which is not rescued by lovastatin ( YWHAG R132C/+ with DMSO = 0.0201 ± 0.000343; YWHAG R132C/+ with lovastatin = 0.0198 ± 0.000511; LME accounting for batch, p=0.98; n=3 independent biological replicates), and the amplitude of YWHAG +/+ calcium spikes remains higher than in YWHAG R132C/+ neurons comparison with ( YWHAG R132C/+ YWHAG +/+ with lovastatin = 0.0224 ± 0.000348, YWHAG R132C/+ with lovastatin = 0.0198 ± 0.000511; LME accounting for batch, p = 0.0018). Colors indicate batches, d) sampling of 20 random calcium traces.

Article Snippet: Neuronal cultures were treated with 10 μM lovastatin (Tocris, 1530) or 10 μM Y27632 (abcam, ab144494) for 1 hour, followed by incubation with Fluo-4 AM dye (Thermo Fisher Scientific, F14201) as described above (a total of 1 hour and 30 minutes of treatment) prior to live cell calcium imaging.

Techniques: Control, Comparison, Sampling

Journal: bioRxiv

Article Title: Decoupled calcium homeostasis and signaling associated with cytoskeletal instability in YWHAG R132C induced pluripotent stem cell-derived cortical neurons

doi: 10.64898/2026.04.01.715876

Figure Lengend Snippet: a) Fold change of YWHAG R132C/+ versus control. N=3 independent biological replicates at d.i.v. 7 and d.i.v. 30, matching the RNAseq timepoints in ). b) Treatment with the ROCK inhibitor Y27632 results in a shift in calcium baseline (stats) c) Western blot quantification of protein expression in YWHAG R132C/+ and isogenic control neurons. Modulation of the ROCK pathway with Y27632 and lovastatin did not alter the protein levels of 14-3-3γ. d) Trypsin-EDTA (0.05%)-induced cytoskeletal destabilization leads to a drastic loss of cytoskeletal proteins and HSP70. This is partially rescued by lovastatin, which restores protein stability. e) Quantification of 14-3-3γ protein levels upon Trypsin-EDTA mediated stress and the lovastatin rescue ( YWHAG +/+ Mean FC lovastatin/DMSO ± S.E.M. = 1.16 ± 0.08, YWHAG R132C/+ = 2.45 ± 0.241; one sample t-test on log2FC lovastatin/DMSO; YWHAG +/+ p=0.161; YWHAG R132C/+ p=0.00395; n=4, independent biological replicates, intensity in arbitrary units).

Article Snippet: Neuronal cultures were treated with 10 μM lovastatin (Tocris, 1530) or 10 μM Y27632 (abcam, ab144494) for 1 hour, followed by incubation with Fluo-4 AM dye (Thermo Fisher Scientific, F14201) as described above (a total of 1 hour and 30 minutes of treatment) prior to live cell calcium imaging.

Techniques: Control, RNA sequencing, Western Blot, Expressing