n2 r d systems ar009  (R&D Systems)

Journal: The EMBO Journal

Article Title: Cellular quiescence uncouples the proteome from the transcriptome in neural stem cells

doi: 10.1038/s44318-026-00693-4

Figure Lengend Snippet: EdU intensity was determined for individual nuclei and normalised to average of active condition. Following BMP4 wash-out, qNSCs that had been exposed to BMP4 for 3 d reactivated faster than those exposed to BMP4 for 10 d. Histograms represent mean and error bars s.e.m. of one of n = 2 biological replicates; phenotype averages normalised to aNSC average were: 3 d BMP4 0.0003; 3 d BMP4 + 1 d washout 0.004; 3 d BMP4 + 2 d washout 0.15; 3 d BMP4 + 3 d washout 0.21; 3 d BMP4 + 4 d washout 0.37; 3 d BMP4 + 5 d washout 0.34; 3 d BMP4 + 6 d washout 0.68; 10 d BMP4 0.005; 10 d BMP4 + 2 d washout 0.007; 10 d BMP4 + 4 d washout 0.006; 10 d BMP4 + 6 d washout 0.006; 10 d BMP4 + 8 d washout 0.006; 10 d BMP4 + 10 d washout 0.006; 10 d BMP4 + 12 d washout 0.06; 10 d BMP4 + 14 d washout 0.14; 10 d BMP4 + 16 d washout 0.30; 10 d BMP4 + 18 d washout 0.50; 10 d BMP4 + 21 d washout 0.80. .

Article Snippet: NSCs were then propagated in Dulbecco’s modified Eagle’s medium (DMEM)/F-12 with Glutamax (Thermo Fischer Scientific 31331093) + N-2 MAX supplement (R&D Systems AR009) + penicillin–streptomycin (Thermo Fischer Scientific 15140) + 2 mg/ml laminin (Sigma L2020) + 5 mg/ml heparin (Sigma H3393-50KU) + 20 ng/ml recombinant murine fibroblast growth factor 2 (FGF-2) (PeproTech 450-33) + 20 ng/ml recombinant murine epidermal growth factor (EGF) (PeproTech 315-09) in tissue culture incubators at 37 °C with 5% CO 2 . qNSC induction was performed with 50 ng/ml BMP4 (R&D Systems 5020-BP-010) and qNSCs cultures were never passaged.

Techniques:

Journal: The EMBO Journal

Article Title: Cellular quiescence uncouples the proteome from the transcriptome in neural stem cells

doi: 10.1038/s44318-026-00693-4

Figure Lengend Snippet: ( A ) Average fraction of transcripts containing five common types of mRNA multivalency (which can coexist in the same transcript) plotted for nuclear biasing, cytoplasmic biasing, or mRNAs not changing subcellular bias from n = 4 biological replicates (fracRNA-seq data). ( B ) Co-staining of oligo(dT) with the nuclear speckle markers Srrm2 and phosphor-SR in aNSCs versus qNSCs of increasing quiescence depth. All scale bars: 10 µm. ( C ) Quantification of oligo(dT) and phosphor-SR speckle/nucleoplasmic signal from individual cells such as those depicted in ( B ) from one of n = 2 biological replicates; white lines: median and quartile boundaries. Median values displayed over dot plots. Phenotype averages were normalised to aNSC and were: oligo(dT) in 3d-qNSC 1.43 and in 10d-qNSC 1.71; phospho-SR in 3d-qNSC 1.30 and in 10d-qNSC 1.98. Mann–Whitney test: **** P < 0.0001. ANOVA test for each: **** P < 0.0001. ( D ) Quantification of Psap, Eif3a and Map1B mRNA speckle/nucleoplasmic signal from individual cells in aNSCs versus qNSCs of increasing quiescence depth (representative images for each transcript can be found in Fig. ) from one of n = 2 biological replicates; white lines: median and quartile boundaries. Median values are displayed over dot plots. Phenotype averages were normalised to aNSC and were: Psap in 3d-qNSC 1.62 and in 10d-qNSC 1.26; Eif3a in 3d-qNSC 2.37 and in 10d-qNSC 2.61; Map1b in 3d-qNSC 2.28 and in 10d-qNSC 1.61. Mann–Whitney test, *** P < 0.001, **** P < 0.0001. ANOVA test for each probe: **** P < 0.0001. ( E ) Model: altered speckle composition in qNSCs, namely with increased SR proteins, turn it into a “sink” for some mRNAs, namely GA-rich multivalent ones, which become nuclear-retained. .

Article Snippet: NSCs were then propagated in Dulbecco’s modified Eagle’s medium (DMEM)/F-12 with Glutamax (Thermo Fischer Scientific 31331093) + N-2 MAX supplement (R&D Systems AR009) + penicillin–streptomycin (Thermo Fischer Scientific 15140) + 2 mg/ml laminin (Sigma L2020) + 5 mg/ml heparin (Sigma H3393-50KU) + 20 ng/ml recombinant murine fibroblast growth factor 2 (FGF-2) (PeproTech 450-33) + 20 ng/ml recombinant murine epidermal growth factor (EGF) (PeproTech 315-09) in tissue culture incubators at 37 °C with 5% CO 2 . qNSC induction was performed with 50 ng/ml BMP4 (R&D Systems 5020-BP-010) and qNSCs cultures were never passaged.

Techniques: Staining, MANN-WHITNEY

Journal: The EMBO Journal

Article Title: Cellular quiescence uncouples the proteome from the transcriptome in neural stem cells

doi: 10.1038/s44318-026-00693-4

Figure Lengend Snippet: ( A – C ) Representative pictures from one of n = 2 biological replicates of the nuclear speckle marker Srrm2, in aNSCs versus qNSCs of increasing quiescence depth co-stained with: ( A ) Psap ; ( B ) Eif3a ; and ( C ) Map1b mRNAs. The final column of each set is the merge of the magenta and yellow channels. Images are MAX intensity projections of Z-stacks. Scale bars: 10 µm. Signal quantifications are presented in Fig. . .

Article Snippet: NSCs were then propagated in Dulbecco’s modified Eagle’s medium (DMEM)/F-12 with Glutamax (Thermo Fischer Scientific 31331093) + N-2 MAX supplement (R&D Systems AR009) + penicillin–streptomycin (Thermo Fischer Scientific 15140) + 2 mg/ml laminin (Sigma L2020) + 5 mg/ml heparin (Sigma H3393-50KU) + 20 ng/ml recombinant murine fibroblast growth factor 2 (FGF-2) (PeproTech 450-33) + 20 ng/ml recombinant murine epidermal growth factor (EGF) (PeproTech 315-09) in tissue culture incubators at 37 °C with 5% CO 2 . qNSC induction was performed with 50 ng/ml BMP4 (R&D Systems 5020-BP-010) and qNSCs cultures were never passaged.

Techniques: Marker, Staining