FISH was performed with minor modifications from the method described in the QuantiGene ViewRNA ISH Cell Assay kit (Affymetrix). Briefly, adherent neuronal cultures were washed in Tyrode’s solution before being fixed in 4% PFA (Sigma) in PBS. Samples were further washed in PBS, and the detergent solution was provided by the manufacturer. Specific neuroLNC or glyceraldehyde phosphate dehydrogenase (GAPDH) oligonucleotide probes were designed and ordered from Affymetrix, diluted in probe diluent solution, and used for the hybridization. Coverslips were incubated upside down on the probe mix solution in a humidified chamber for 3 hours at 40°C. Alternating with washes in SSC buffer (UltraPure SSC, Thermo Fisher Scientific), incubation with the preamplifier, amplifier, and label mix (1:50 in respective diluent; 30 min at 40°C each) followed. Nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole), and coverslips were embedded on object slides in Mowiol (Calbiochem, Merck).

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