Fig. 8

TNF-α increases the expression of Nav1.6 in the retina. a–c Bar charts show the relative mRNA levels of Nav1.1, Nav1.2, and Nav1.6 in Ctr (a), TNF-α3D (b), and TNF-α7D (c) groups. n = 5 for each group. d Representative immunoblots show the changes in total Nav1.6 (tNav1.6) expression in control (Ctr), TNF-α3D, TNF-α7D, and COH2W without or with R7050 retinal extracts. e Bar charts summarize the average densitometric quantification of immunoreactive bands of tNav1.6 under different conditions as shown in d. n = 6 for each group. All data are normalized to their corresponding β-actin and then to Ctr. f Representative immunoblots show the changes in Nav1.6 expression in membrane components (mNav1.6) in control (Ctr), TNF-α3D, TNF-α7D, and COH2W retinal extracts. n = 4 for each group. All data are normalized to their corresponding GAPDH and then to Ctr. *P < 0.05 and ***P < 0.001 vs. Ctr. h Immunofluorescence labeling showing Nav1.6 expression in rat retinal vertical slices taken from the normal saline-injected retina (Ctr, h1-h3) and TNF-α-injected retinas on 7 days after the injection (TNF-α7D, h4-h6). h7-h9: double immunofluorescence staining showing Nav1.6 expression when the Nav1.6 antibody was pre-absorbed with its blocking peptide (BP). Note that the enhanced expression of Nav1.6 in the axon-like structures in the TNF-α7D group. Scale bars = 20 μm. GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer