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Fig. 3 | Acta Neuropathologica Communications

Fig. 3

From: The involvement of tau in nucleolar transcription and the stress response

Fig. 3

Nucleolar stress co-occurs with the redistribution of nucleolar nP-Tau. a Flow cytometry experiments with CellROX Green following 20 mM Glutamate treatment of differentiated SHSY5Y showed oxidative stress [P = 0.0013]. b Western blotting analysis revealed that the glutamate treatment led to a significant decrease in TIP5, UBF, and FBL. [TIP5 P < 0.0001]; [UBF P = 0.0004]; [FBL P = 0.0002]. c qPCR analysis of rDNA transcription and processing showed that the glutamate incubation resulted in a significant decrease in 45S pre-rRNA synthesis [45S pre-rRNA P = 0.008]. d Representative immunofluorescence fluorescence images showing labelling for nP-Tau and FBL control and following glutamate treatment (Arrows showing regions in which colocalisation of nP-Tau and FBL is altered by Glutamate treatment). Graphs showing quantification from four independent experiments each with five images and each containing an average of 35 cells. Glutamate administration resulted in redistribution of nucleolar nP-Tau from FBL (blue arrow), as well as FBL redistribution from nP-Tau (white arrows) compared to the control. Analysis of immunofluorescence reveals a significant increase in the number (33%) of cells showing FBL redistribution (dii) [P < 0.02]. Quantification revealed that 14% of Glutamate-treated cells showed nucleolar nP-Tau redistribution (diii). [P < 0.02]. Total level of nuclear nP-Tau is increased (div) [P < 0.001]. dv Western blotting on whole cell extracts showed a significant increase in nP-Tau, with no changes in T-Tau levels. nP-Tau [P < 0.0001]; T-Tau: [P = 0.47]. Intensity normalised to β-actin. Images showing nucleolar tau and FBL in untreated and treated cells were Z-projected for maximum intensity. For all experiments N ≥ 4

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