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. 2020 Oct 20;28(3):1041–1061. doi: 10.1038/s41418-020-00636-4

Fig. 7. Exosomal counteraction of N-methyl-N-nitrosourea (MNU) damages on photoreceptor cells involves inhibition of the miR-21 target programmed cell death 4 (Pdcd4).

Fig. 7

Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of mRNA expression levels of multiple targets of miR-21 in retina tissues (A) and 661W cone photoreceptor cells (B), normalized to Glyceraldehyde-3-phosphate dehydrogenase (Gapdh). Ctrl control, Spry1 Sprouty 1, Spry 2 Sprouty 2, Tfap2a Transcription factor AP-2-alpha. *P < 0.05 by the Mann–Whitney U tests. N = 4 per group. qRT-PCR analysis of mRNA expression levels (C) and western blot analysis of protein expression levels (D) with corresponding quantification (E) of Pdcd4 in retina tissues, normalized to Gapdh. EXOT exosomal transplantation after MNU injection. *P < 0.05 by the Kruskal–Wallis tests. N = 3 per group. qRT-PCR analysis of mRNA expression levels (F) and western blot analysis of protein expression levels (G) with corresponding quantification (H) of Pdcd4 in 661W cells, normalized to Gapdh. *P < 0.05 by the Kruskal–Wallis tests. N = 3 per group. Western blot analysis of protein expression levels (I) with corresponding quantification (J) of Pdcd4 in 661W cells, normalized to Gapdh. NC negative control of miR-21 mimics. 661W cells were treated with MNU with or without transfection of NC or miR-21 mimics. *P < 0.05 by the Kruskal–Wallis test. N = 3 per group. Western blot analysis of protein expression levels (K) with corresponding quantification (L) of Pdcd4 in 661W cells, normalized to Gapdh. siRNA-NC negative control of small interfering RNA for Pdcd4, siPdcd4 small interfering RNA for Pdcd4. 661W cells were treated with MNU with or without transfection of siRNA-NC or siPdcd4. *P < 0.05 by the Kruskal–Wallis test. N = 3 per group. Representative flow cytometric images showing death of 661W cone photoreceptor cells (M) and the corresponding quantitative analysis of percentages of apoptotic (Annexin V+PI− plus Annexin V+PI+) 661W cells (N). 661W cells were treated with MNU with or without transfection of siRNA-NC or siPdcd4. *P < 0.05 by the Kruskal–Wallis test. N = 4 per group. Data represent median ± range for (C), (E), (F), (H), (J), and (L). Data are represented as box (25th, 50th, and 75th percentiles) and whisker (range) plots for (A), (B), and (N).