Journal: Heliyon

Article Title: CD133-containing microvesicles promote colorectal cancer progression by inducing tumor angiogenesis

doi: 10.1016/j.heliyon.2024.e29292

Figure Lengend Snippet: CD133-containing MVs are incorporated into endothelial cells. (A) Stable shMock and shCD133 HCT116 cells were generated. MVs were isolated from shMock and shCD133 HCT116 cells. Protein level was determined using western blotting. Full-length blot images are shown in (B) shMock and shCD133 MVs were stained with WGA-488. EA.hy926 cells were incubated with stained MVs for 16 h. Images were captured by confocal laser scanning microscopy (green, WGA-488; blue, DAPI). Scale bar = 10 μm. The number of MVs were counted by imageJ. All p values were obtained using unpaired two-tailed Student's t -test. *p < 0.05. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Mouse monoclonal antibodies recognizing CD133 were purchased from the Developmental Studies Hybridoma Bank (Iowa City, IA, USA).

Techniques: Generated, Isolation, Western Blot, Staining, Incubation, Confocal Laser Scanning Microscopy, Two Tailed Test

Journal: Heliyon

Article Title: CD133-containing microvesicles promote colorectal cancer progression by inducing tumor angiogenesis

doi: 10.1016/j.heliyon.2024.e29292

Figure Lengend Snippet: CD133-containing MVs promote tube formation and vessel sprouting in endothelial cells. (A) EA.hy926 cells were treated with MVs isolated from shMock and shCD133 HCT116 cells for 16 h. Scale bar = 100 μm. (B) Meshes and tubes were measured from five random fields in each well. (C) EA.hy926 cell spheroids embedded in 3D collagen gels were treated with shMock or shCD133 MVs for 48 h. Representative images of endothelial spheroids were shown. Scale bar = 100 μm. (D) Five random spheroids were measured in each well. (E) Mouse aortic rings were embedded in 3D collagen gels and stimulated with 2% FBS media supplemented with VEGF (30 ng/mL). After 48 h, rings were washed twice, and media were changed with 2% FBS containing shMock or shCD133 MVs and incubated for 3 days. Representative images of mouse aortic rings were shown. Scale bar = 200 μm. (F) All the sprouts of each ring were measured. Mean ± SEM of n = 3 independent experiments. All p values were obtained using unpaired two-tailed Student's t -test. *p < 0.05, * *p < 0.01 ** *p < 0.001.

Article Snippet: Mouse monoclonal antibodies recognizing CD133 were purchased from the Developmental Studies Hybridoma Bank (Iowa City, IA, USA).

Techniques: Isolation, Incubation, Two Tailed Test

Journal: Heliyon

Article Title: CD133-containing microvesicles promote colorectal cancer progression by inducing tumor angiogenesis

doi: 10.1016/j.heliyon.2024.e29292

Figure Lengend Snippet: CD133-containing MVs induce angiogenesis through the p38 MAPK signaling pathway. (A) EA.hy926 cells were treated with shMock and shCD133 MVs for 16 h. Protein level was determined using western blotting. p-p38, phosphorylated p38; pAKT, phosphorylated AKT; pERK, phosphorylated ERK. Full-length blot images are shown in (B) Quantification of the p-p38 level. (C) EA.hy926 cells were pre-treated with SB203580 (20 μM) for 2 h and treated with shMock MVs in the presence of SB203580 for 13 h. Representative images of tube formation were shown. Scale bar = 100 μm. (D) Meshes and tubes were measured from five random fields in each well. Mean ± SEM of n = 3 independent experiments. All p values were obtained using an unpaired two-tailed Student's t -test. *p < 0.05, * *p < 0.01, ** *p < 0.001.

Article Snippet: Mouse monoclonal antibodies recognizing CD133 were purchased from the Developmental Studies Hybridoma Bank (Iowa City, IA, USA).

Techniques: Western Blot, Two Tailed Test

Journal: Heliyon

Article Title: CD133-containing microvesicles promote colorectal cancer progression by inducing tumor angiogenesis

doi: 10.1016/j.heliyon.2024.e29292

Figure Lengend Snippet: CD133-containing MV-induced angiogenesis is not mediated by the canonical angiogenesis pathways. (A) Angiogenesis antibody array was performed using EA.hy926 cells treated with shMock MVs or shCD133 MVs. Red represents high protein expression levels, and blue represents low protein expression levels. (B) Protein expression was determined using western blot analysis. Human recombinant VEGF165 protein was used as a control. Full-length blot images are shown in . (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Mouse monoclonal antibodies recognizing CD133 were purchased from the Developmental Studies Hybridoma Bank (Iowa City, IA, USA).

Techniques: Ab Array, Expressing, Western Blot, Recombinant

Journal: Heliyon

Article Title: CD133-containing microvesicles promote colorectal cancer progression by inducing tumor angiogenesis

doi: 10.1016/j.heliyon.2024.e29292

Figure Lengend Snippet: Angiogenesis induced by CD133-containing MVs is resistant to bevacizumab and CD133 overexpression predicts poor prognosis in metastatic CRC patients. (A) EA.hy926 cells were treated with VEGF (200 ng/mL) or shMock MVs in the presence or absence of bevacizumab (100 μg/mL) for 16 h. Representative images of tube formation. Scale bar = 100 μm. (B) Meshes and tubes were measured from five random fields in each well. Mean ± SEM of n = 3 independent experiments. All p values were obtained using an unpaired two-tailed Student's t -test. *p < 0.05, * *p < 0.01, ** *p < 0.001. (C) The correlation between CD133 mRNA expression level and patient survival rate was analyzed. Kaplan-Meier plots were obtained from the Human Protein Atlas database. The metastatic CRC survival rate represents patients in stages Ⅲ and Ⅳ. The overall survival rate represents CRC patients in all stages.

Article Snippet: Mouse monoclonal antibodies recognizing CD133 were purchased from the Developmental Studies Hybridoma Bank (Iowa City, IA, USA).

Techniques: Over Expression, Two Tailed Test, Expressing

Journal: Heliyon

Article Title: CD133-containing microvesicles promote colorectal cancer progression by inducing tumor angiogenesis

doi: 10.1016/j.heliyon.2024.e29292

Figure Lengend Snippet: Schematic diagram regarding the role of CD133-containing MVs derived from CRC cells in angiogenesis and CRC progression.

Article Snippet: Mouse monoclonal antibodies recognizing CD133 were purchased from the Developmental Studies Hybridoma Bank (Iowa City, IA, USA).

Techniques: Derivative Assay

Journal: PeerJ

Article Title: DNA mismatch repair and CD133-marked cancer stem cells in colorectal carcinoma

doi: 10.7717/peerj.5530

Figure Lengend Snippet: (A) The weak CD133 staining seen at the luminal border of some normal colonic crypts is considered equivocal and interpreted as negative (x200). In contrast, (B) is a case of colorectal carcinoma with strong CD133 staining of the glandular luminal surface (x200) and (C) a case of poorly differentiated colorectal carcinoma demonstrating CD133 staining as tiny cytoplasmic blobs which represent abortive glands (x200). (D) Colorectal carcinoma with deficient DNA mismatch repair (represented by loss of MLH1). Note the lack of immunoexpression of MLH1 in the colorectal carcinoma nuclei in the presence of immunopositivity in the internal positive controls (stromal cells, lymphocytes and nearby normal enterocytes) (x200)

Article Snippet: Immunohistochemical staining with a rabbit polyclonal antibody to CD133 (1:200; Abcam ab19898, Cambridge, United Kingdom) and detection by the ultraView universal DAB detection kit (Ventana Medical Systems Inc., Tucson, AZ, USA) was carried out on the Ventana Benchmark XT autostainer (Ventana Medical Systems Inc., Tucson, AZ, USA).

Techniques: Staining

Journal: PeerJ

Article Title: DNA mismatch repair and CD133-marked cancer stem cells in colorectal carcinoma

doi: 10.7717/peerj.5530

Figure Lengend Snippet: CD133 expression and DNA mismatch repair (MMR) status. The CD133 expression and MMR status as per size, TNM stage and differentiation of the colorectal carcinoma ( n = 80).

Article Snippet: Immunohistochemical staining with a rabbit polyclonal antibody to CD133 (1:200; Abcam ab19898, Cambridge, United Kingdom) and detection by the ultraView universal DAB detection kit (Ventana Medical Systems Inc., Tucson, AZ, USA) was carried out on the Ventana Benchmark XT autostainer (Ventana Medical Systems Inc., Tucson, AZ, USA).

Techniques: Expressing

Journal: PeerJ

Article Title: DNA mismatch repair and CD133-marked cancer stem cells in colorectal carcinoma

doi: 10.7717/peerj.5530

Figure Lengend Snippet: DNA mismatch repair protein (MMR) status and CD133 expression. CD133 immunoexpression and score being stratified by MMR status in right-sided and left-sided colorectal carcinomas.

Article Snippet: Immunohistochemical staining with a rabbit polyclonal antibody to CD133 (1:200; Abcam ab19898, Cambridge, United Kingdom) and detection by the ultraView universal DAB detection kit (Ventana Medical Systems Inc., Tucson, AZ, USA) was carried out on the Ventana Benchmark XT autostainer (Ventana Medical Systems Inc., Tucson, AZ, USA).

Techniques: Expressing