Figure 1.

The photomicrographs depict the immunohistochemical labeling of voltage-gated sodium channels (VGSCs) (green) in a biopsy of the stage IIB carcinoma of the cervix obtained from the patient prior to treatment with targeted osmotic lysis (TOL). Sections were incubated and then processed with a goat-anti-rabbit 488 Alexa fluorophore secondary antibody, 1:800 dilution. Nuclei were counterstained with DRAQ5 (blue; dilution 1:600). Twelve images taken with a Leica DMi8 confocal microscope at 40× oil magnification were reviewed for evidence of VGSC labeling. Panel (A) depicts a representative portion of the biopsy where many cells, approximately 40% of the total, highly expressed VGSCs. Cellular profiles from three sections were outlined and the mean fluorescence intensity was measured using the ImageJ-FIJI software (imagej.nih.gov; accessed on 19 May 2021) to quantify the sodium channel expression. The histogram (B) presents the average mean fluorescence intensity measured from 50 intensely stained and 50 weakly stained cell profiles compared to the background indicative of the level of VGSC expression in highly expressing tumor cells and minimally expressing neoplastic and stromal cells, respectively. A one-way ANOVA was significant (p < 0.00001); * = p < 0.001 compared to the background; ** = p < 0.001 compared to weakly expressing cells (Student’s t-test with Scheffe’s adjustment). Calibration bar in A = 25 µm.