Journal: Journal of Extracellular Biology

Article Title: Chemo‐small extracellular vesicles released in cisplatin‐resistance ovarian cancer cells are regulated by the lysosomal function

doi: 10.1002/jex2.157

Figure Lengend Snippet: A2780cis CDDP‐resistant cells have more capacity to produce and secrete exosomes. (a) Representative confocal microscopy images of A2780 and A2780cis PFA‐fixed cells immunofluorescent stained with anti‐CD63. Scale Bar 10 μm. (b) Analysis of the number of structures and (c) area of structures of CD63 per cell from images as those shown in (a). Bars indicated the mean with SEM; *** p < 0.001; one‐tiled unpaired parametric t ‐test ( n > 50 cells from three independent experiments). (d) Representative TEM micrograph shows MVEs of A2780 and A2780cis cells. Scale Bar 0.5 μm. (e) Semiquantitative analysis of the area of MVEs and (f) ILVs per MVEs of A2780 and A2780cis cells, from images as those shown in (d). Bars indicated the mean with SEM; * p < 0.05, *** p < 0.001; one‐tiled, unpaired, parametric t ‐test ( n > 50 cells from three independent experiments). (g) Analysis of detergent‐soluble protein extracts obtained from A2780 and A2780cis cells by Western blot with anti‐HRS, anti‐ALIX, anti‐TSG101, anti‐LAMP2A and anti‐β‐actin. The image is representative of three independent experiments. (h) Densitometric quantification of the signal of HRS, ALIX, TSG101 and LAMP2A from images as those shown in (g). The signal was normalized with β ‐actin signal. Bars indicated the mean with SEM; * p < 0.05, ** p < 0.01, one‐tiled, paired, non‐parametric Mann‐Whitney test ( n = 3). (i) Analysis of detergent‐soluble protein extracts obtained from A2780 and A2780cis cells by Western blot with anti‐RAB11A, anti‐RAB22A, anti‐RAB27A, anti‐RAB35 and anti‐β‐actin. The image is representative of three independent experiments. (j) Densitometric quantification of the signal of RAB11A, RAB22A, RAB27A and RAB35 from images as those shown in (i). The signal was normalized with β ‐actin signal. Bars indicated the mean with SEM; ** p < 0.01, NS: not significant; one‐tiled, paired non‐parametric Mann‐Whitney test ( n = 3).

Article Snippet: In general, the type and quantity of EVs are controlled intracellularly through its biogenesis at the level of MVEs and ILVs and/or with the trafficking and fusion of MVEs with the PM (Dixson et al., ; Steinbichler et al., ; van Niel et al., ).

Techniques: Confocal Microscopy, Staining, Western Blot, MANN-WHITNEY