Journal: Oncology Reports

Article Title: Exosomes derived from retinoblastoma cells enhance tumour deterioration by infiltrating the microenvironment

doi: 10.3892/or.2020.7858

Figure Lengend Snippet: EXOs derived from WERI-Rb1 cells could promote the malignancy of retinoblastoma. (A) Vimentin (green) and Ki-67 (red) were labelled by immunofluorescence. Scale bar, 150 µm. (B) Quantitative data showing the relative positive area of Vimentin and Ki-67 in tumours (n=5). (C) Western blotting showed CXCR4 and MMP2 protein expression in tumours from the PBS-injected and EXO-injected groups. (D) Semi-quantification of CXCR4 and MMP2 expression in tumours (n=5). (E) Staining of CD68 (green) in PBS-injected and EXO-injected tumours. Scale bar, 2 mm or 100 µm. (F) Quantitative data showed that CD68-positive cells were increased by EXO injection (n=6). (G) Staining of CD49b (red) in PBS-injected and EXO-injected tumours. Scale bar, 50 µm. (H) Quantitative data showed that CD49b-positive cells were decreased (n=5). *P<0.05 and ***P<0.001. EXO, exosome; CXCR4, C-X-C chemokine receptor type 4.

Article Snippet: Samples were incubated with primary antibodies overnight at 4°C, including anti-CD49b for natural killer (NK) cells (1:100; cat. no. 14-5971-85; Invitrogen; Thermo Fisher Scientific, Inc.), anti-CD45 for leukocytes (1:100; cat. no. sc-1178; Santa Cruz Biotechnology, Inc.), anti-CD68 for macrophages (1:100; cat. no. BA3638; Wuhan Boster Biological Technology Co., Ltd.), anti-Ki-67 for proliferative cells (1:500; cat. no. MA5-14520; Thermo Fisher Scientific, Inc.), anti-Vimentin for invasive tumour cells (1:100; cat. no. sc-6260; Santa Cruz Biotechnology, Inc.).

Techniques: Derivative Assay, Immunofluorescence, Western Blot, Expressing, Injection, Staining

Journal: Oncology Reports

Article Title: Exosomes derived from retinoblastoma cells enhance tumour deterioration by infiltrating the microenvironment

doi: 10.3892/or.2020.7858

Figure Lengend Snippet: EXOs derived from WERI-Rb1 cells inhibit innate immunity in vivo . PKH26 EXOs (red) were internalized by cells from spleen tissues (A) 2 or (B) 10 days following EXO injection. Scale bar, 50 or 20 µm. (C) EXOs labelled by PKH26 were engulfed by CD68-positive macrophages in the spleen (green, CD68; red, PKH26; blue, DAPI). Scale bar, 5 µm. (D) Ki-67 (red) and CD45 (green) were labelled by immunofluorescence in spleens from the PBS-injected or EXO-injection groups. Scale bar, 200 or 50 µm. (E) Quantification data showed CD45-positive cells in spleens from the PBS-injected and EXO-injected groups (n=5). (F) Quantification of CD45 and Ki-67 double-positive cells in spleens from the PBS-injected and EXO-injected groups (n=5). (G) Staining of CD49b (red) in spleens from the Sham (normal mice), PBS-injected and EXO-injected groups. Scale bar, 50 µm. (H) Quantification of CD49b-positive cells in spleens from the Sham (n=3), PBS-injected (n=5) and EXO-injected groups (n=5). *P<0.05, **P<0.01 and ***P<0.001. EXO, exosome.

Article Snippet: Samples were incubated with primary antibodies overnight at 4°C, including anti-CD49b for natural killer (NK) cells (1:100; cat. no. 14-5971-85; Invitrogen; Thermo Fisher Scientific, Inc.), anti-CD45 for leukocytes (1:100; cat. no. sc-1178; Santa Cruz Biotechnology, Inc.), anti-CD68 for macrophages (1:100; cat. no. BA3638; Wuhan Boster Biological Technology Co., Ltd.), anti-Ki-67 for proliferative cells (1:500; cat. no. MA5-14520; Thermo Fisher Scientific, Inc.), anti-Vimentin for invasive tumour cells (1:100; cat. no. sc-6260; Santa Cruz Biotechnology, Inc.).

Techniques: Derivative Assay, In Vivo, Injection, Immunofluorescence, Staining

Journal: Oncology Reports

Article Title: Exosomes derived from retinoblastoma cells enhance tumour deterioration by infiltrating the microenvironment

doi: 10.3892/or.2020.7858

Figure Lengend Snippet: EXOs derived from WERI-Rb1 cells accelerate retinoblastoma metastasis. PKH26-labelled EXOs (red) were internalized by CD68-positive Kupffer cells (green) from liver tissues (A) 2 or (B) 10 days following EXO injection. DAPI was used to stain nuclei, and CD68 (green) was used to label macrophages. Scale bar, 50 or 10 µm. (C) Macroscopic appearance of livers from the PBS-injected and EXO-injected group (white arrowhead, metastatic liver nodules; green arrowheads, liver cirrhosis). (D) Staining of Ki-67 (red) in livers from the PBS-injected and EXO-injected groups. Scale bar, 200 µm. (E) Quantitative data showed that the Ki-67-positive area (%) was increased by EXO injection (n=5). **P<0.01. EXO, exosome.

Article Snippet: Samples were incubated with primary antibodies overnight at 4°C, including anti-CD49b for natural killer (NK) cells (1:100; cat. no. 14-5971-85; Invitrogen; Thermo Fisher Scientific, Inc.), anti-CD45 for leukocytes (1:100; cat. no. sc-1178; Santa Cruz Biotechnology, Inc.), anti-CD68 for macrophages (1:100; cat. no. BA3638; Wuhan Boster Biological Technology Co., Ltd.), anti-Ki-67 for proliferative cells (1:500; cat. no. MA5-14520; Thermo Fisher Scientific, Inc.), anti-Vimentin for invasive tumour cells (1:100; cat. no. sc-6260; Santa Cruz Biotechnology, Inc.).

Techniques: Derivative Assay, Injection, Staining