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. 2015 Jul 27;5:12425. doi: 10.1038/srep12425

Figure 4. BA desastibilized lysosomes of HaCaT keratinocytes.

Figure 4

(a) After treatment for 6 hours with DMSO (control), triterpenoids (20 μM) or TEM (15 μM) cells were stained with LTG and evaluated under confocal microscope. Surface plots below the images represented the fluorescence of LTG-loaded lysosomes. (b) Within 6 hours of DMSO (control) and triterpenoids (20 μM) addition, cells were stained with LTR (red) and immunostained for total CTSB (green). (i) Fluorescence plot profiles represented line scans of LTR-loaded lysosomes and CTSB (bottom). Micrographs of BA-treated cells (lined area) showing elevated CTSB in cytosol associated with decrease in LTR-loaded lysosomes (ii). At right, surface plots of depicted area drawn in the images (ii). (c) Cathepsin B (CTSB) activity from cytosolic fraction within 3 h of BA (20 μM) addition upon absence (–) or presence of CTSB inhibitor CA074 at 5 μM (+) and 10 μM (++). T1 = after treatment for 24 hours; T2 = 24 hours after T1; T3 = 48 h after T1. All results were obtained from at least three independent experiments and expressed as mean values ± standard error. Multiple statistical comparisons were calculated by ANOVA test, and the p-value for each pairwise group was determined by Dunnett’s T3 (high variance between groups) or Bonferroni (low variance between groups) post-hoc test. Significance difference (p < 0.05) was depicted by asterisk.