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. 2016 Sep 9;6:32960. doi: 10.1038/srep32960

Figure 6. Expression of HMGB4 and HMGB4L1 in cultured primary cells of rat brain.

Figure 6

(a) Immunofluorescence staining of nestin and HMGB4L1 in 1d differentiated rat neuronal cells in vitro. Neurospheres were allowed to adhere and differentiate for 1 day and cells were immunofluorescently stained with anti-nestin and anti-HMGB4L1 antibodies. Cell nuclei were stained with DAPI. The staining controls for anti-nestin or anti-HMGB4L1 staining were done without primary antibody or with preimmune serum, respectively. Scale bar = 30 μm. (b) Immunofluorescence staining of HMGB4L1 and NeuN in 14d differentiated rat neuronal cells in vitro. Neurospheres were allowed to differentiate for 14 days and cells were immunofluorescently stained with anti-NeuN and anti-HMGB4L1 antibodies. Staining controls for anti-HMGB4L1 and anti-NeuN were done with pre-immune serum or without primary antibody, respectively. Scale bar = 20 μm. (c,d) Proximity ligation assay of neurospheres with anti-PAN-Histone and anti-HMGB4L1 antibodies. Cell nuclei of cultured neurospheres were stained with DAPI and a proximity ligation assay was performed with anti-PAN-Histone antibodies and preimmune control serum (c) or with anti-PAN-Histone antibodies and anti-HMGB4L1 antibodies (d). Scale bar = 20 μm.