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Figure 8


Fig. 8. FISH and immunohistochemistry for NF-M. (A,A') NF-M FISH of single confocal optical sections from opposite sides of spinal cord or unilaterally injected hnRNP K MO1 tadpole (stage 39/40). Broken outlines surround individual neurons. (B1-D4) Cells from dissociated cell culture. (B1-B3) Control MO neuron stained for NF-M protein (B1), RNA (B2) and RNA/DAPI merged (B3). (C1,C2) Adjacent cells from a unilaterally injected hnRNP K MO culture, viewed for NF-M protein (C1) and RNA (C2). Arrow indicates normal staining for both protein and RNA; arrowhead indicates a cell with no protein and FISH pattern typical of hnRNP K MO-injected spinal cord neurons from whole mount. (D1-D4) Neurons from a bilaterally injected hnRNP K MO culture stained for protein (D1), RNA (D2), DAPI (D3) and RNA/DAPI merged (D4). Scale bars in B1,C1 and D2 apply to B1-3,C1-2 and D1-4, respectively. Bar graph shows qRT-PCR of nuclear and cytoplasmic fractions for NF-M and peripherin RNAs from bilaterally injected hnRNP K MO and control animals assayed at stage 29/30. {Delta}CT, mean (±s.d.) difference in the number of PCR cycles to reach threshold (see text). *{Delta}CT was significantly less for NF-M RNA in hnRNP K MO embryos than for other groups (P<0.01, one-way ANOVA).





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