Journal: Oncology Letters

Article Title: ZBTB20 suppresses tumor growth in glioblastoma through activating the TET1/FAS/caspase‑3 pathway

doi: 10.3892/ol.2024.14491

Figure Lengend Snippet: ZBTB20 promotes apoptosis in glioblastoma cells. (A) Western blotting was used to detect BCL2, BAX, CASP3 and cleaved CASP3 expression in ZBTB20-overexpressing U251 cells, (B) the levels of which were semi-quantified and are presented as histograms. The independent experiment was repeated three times. Data are presented as the mean ± standard deviation. *P<0.05. BCL2, B-cell lymphoma-2; BAX, BCL2 associated X; CASP3, caspase-3; NC, negative control; ZBTB20, zinc finger and BTB domain containing 20.

Article Snippet: Next, the membranes were separately incubated overnight at 4°C with the following antibodies: Polyclonal rabbit anti-human ZBTB20 (dilution, 1:2,000; cat. no. ab127702; Abcam), polyclonal rabbit anti-TET1 (1:1,000; cat. no. A21766; ABclonal Biotech Co., Ltd.), polyclonal rabbit anti-human FAS (1:1,000; cat. no. A2639; ABclonal Biotech Co., Ltd.), polyclonal rabbit anti-human B-cell lymphoma-2 (BCL2; 1:2,000; cat. no. BA0412; Wuhan Boster Biological Technology, Ltd.), polyclonal rabbit anti-human BCL2 associated X (BAX; 1:2,000, cat. no. BA0315-2; Wuhan Boster Biological Technology, Ltd.), monoclonal rabbit anti-human CASP3 (1:1,000, cat. no. AF1213; Beyotime Institute of Biotechnology), monoclonal rabbit anti-human cleaved CASP3 (1:1,000; cat. no. AF1150; Beyotime Institute of Biotechnology), monoclonal rabbit anti-human phosphorylated-extracellular signal regulated kinase (p-ERK; 1:1,000; cat. no. AP0485; ABclonal Biotech Co., Ltd.), polyclonal rabbit anti-human ERK (1:1,000; cat. no. A16686; ABclonal Biotech Co., Ltd.), monoclonal rabbit anti-human p-cyclic AMP-responsive-element-binding protein (CREB; 1:500; cat. no. ab32096; Abcam), monoclonal mouse anti-human CREB (1:500; cat. no. ab178322; Abcam) and monoclonal rabbit anti-human β-actin (1:10,000; cat. no. AC026; ABclonal Biotech Co., Ltd.).

Techniques: Western Blot, Expressing, Standard Deviation, Negative Control

Journal: Scientific Reports

Article Title: Evolutionary conservation of a regulative pathway of erythropoiesis in Poikilothermic vertebrates

doi: 10.1038/s41598-022-06617-6

Figure Lengend Snippet: Light microscopy images of red blood cells from Torpedo marmorata Risso and Caretta caretta . ( a ) Bcl-2 protein is located in the cytoplasm of torpedo basophilic erythroblast. Scale bar as in e . ( b ) In the torpedo acidophilic erythroblast and mature erythrocyte, Bcl-2 and mitochondria have perinuclear position. Scale bar as in e . ( c ) Bcl-X L is slightly detected in the torpedo acidophilic erythroblast (black arrowhead), but not in the mature erythrocyte (white arrowhead). Scale bar: 15 µm. ( d , e ) Bax immunolabeling is shown in torpedo acidophilic erythroblast ( d ) and mature erythrocyte ( e ). No Bax immunolabeling was showing in basophilic erythroblasts. Scale bar: 15.4 µm. ( f ) Demonstration of Bcl-2 immunopositivity in the turtle basophilic erythroblast (black arrowhead) and in the reticulocyte (white arrowhead). The reactivity is seen to decrease in the acidophilic erythroblast and the mature erythrocyte. Scale bar: 12 µm. ( g ) Bcl-X L is detectable in the acidophilic erythroblasts. Scale bar: 12 µm. ( h ) Detail of panel of G showing the intense Bcl-X L positivity of an acidophilic erythroblast, near a negative mature erythrocyte. Scale bar: 17 µm. ( i ) Bax is absent in immature turtle stages, while in mature stages it is localized in the cytoplasm. Scale bar: 15 µm.

Article Snippet: The same procedure was followed for Bcl-X L and Bax immunostaining using rabbit polyclonal anti-Bcl-X L and anti-Bax antibodies (Santa Cruz Biotechnology, Santa Cruz, CA; diluted 1:100 in NGS).

Techniques: Light Microscopy, Immunolabeling

Journal: Scientific Reports

Article Title: Evolutionary conservation of a regulative pathway of erythropoiesis in Poikilothermic vertebrates

doi: 10.1038/s41598-022-06617-6

Figure Lengend Snippet: Light microscopy images of red blood cells from Torpedo after 40 Gy irradiation and after 90 Gy irradiation. ( a ) After 40 Gy irradiation we observed a weakly Bcl-2 immunopositivity in basophilic erythroblasts (white arrowhead) and in acidophilic erythroblast (black arrowhead). ( b ) After 90 Gy irradiation, no Bcl-2 immunopositivity was detected in all three stages of maturation. ( c ) After 40 Gy irradiation, Bcl-X L immunopositivity was detected in the acidophilic erythroblast (black arrowhead). ( d ) After 90 Gy irradiation, a weakly Bcl-X L immunopositivity was observed. ( e ) After 40 Gy irradiation, no Bax immunopositivity was observed. ( f ) After 90 Gy irradiation, Bax was identified in the acidophilic erythroblast (black arrowhead) and in the mature erythrocyte (white arrowhead). In the box, a young erythrocyte displaying Bax positivity in mitochondria. Scale bar: ( a ) 15.5 µm, ( b , f ) 9 µm, ( c , e ) 11.5 µm, ( d ) 15 µm.

Article Snippet: The same procedure was followed for Bcl-X L and Bax immunostaining using rabbit polyclonal anti-Bcl-X L and anti-Bax antibodies (Santa Cruz Biotechnology, Santa Cruz, CA; diluted 1:100 in NGS).

Techniques: Light Microscopy, Irradiation

Journal: Scientific Reports

Article Title: Evolutionary conservation of a regulative pathway of erythropoiesis in Poikilothermic vertebrates

doi: 10.1038/s41598-022-06617-6

Figure Lengend Snippet: Electron microscopy images of red blood cells from Torpedo. ( a ) Immunoelectron microscopy reveals the Bcl-2 positivity, as indicated by colloidal gold particles (10 µm diameter), in the mitochondrial matrix (white arrowhead) and nuclear membrane (black arrowhead) of erythroblasts. ( b ) After 90 Gy irradiation, the localization of the Bcl-2 protein remains unchanged and its expression appears slightly reduced. ( c ) Bcl-X L positivity was detectable by numerous and dispersed gold colloidal particles in cytosol of mature red blood cells. ( d ) Bcl-X L positivity: some gold particle aggregates appears to be dispersed in cytosol after 90 Gy irradiation. ( e ) Weak Bax reactivity is identified in the cytosol of mature erythrocytes. ( f ) An increased expression of Bax was observed in mitochondria and in cytosol after 90 Gy X ray in erythrocytes. Scale bar: 0.5 µm. M mitochondria; N nucleus.

Article Snippet: The same procedure was followed for Bcl-X L and Bax immunostaining using rabbit polyclonal anti-Bcl-X L and anti-Bax antibodies (Santa Cruz Biotechnology, Santa Cruz, CA; diluted 1:100 in NGS).

Techniques: Electron Microscopy, Immuno-Electron Microscopy, Irradiation, Expressing

Journal: Scientific Reports

Article Title: Evolutionary conservation of a regulative pathway of erythropoiesis in Poikilothermic vertebrates

doi: 10.1038/s41598-022-06617-6

Figure Lengend Snippet: Electron microscopy images of mitochondria of Caretta caretta and of Torpedo marmorata Risso. ( a ) Loggerhead erythroblast immunolabeling with Bcl-2 antibody shows the location of Bcl-2 (25 nm colloidal gold) on the nuclear membrane (white arrowhead), cristae of the inner mitochondrial membrane and in the cytosol. ( b ) Bcl-X L immunopositivity (25 nm colloidal gold) is observed on the internal mitochondrial membrane (white arrowhead) of young erythrocyte. ( c ) Bax is detected on the crest of a mitochondrion (white arrowhead) in mature erythrocyte. Scale bar: 0.5 µm. ( d , e ) Illustrations of Bcl-2 immunopositivity located on the internal mitochondrial membrane of torpedo young erythrocyte. ( f , g ) Bax positivity is observed to be mainly dispersed on the internal crests of the mitochondria of mature erythrocyte. Scale bar: 0.3 µm. M mitochondria, N nucleus.

Article Snippet: The same procedure was followed for Bcl-X L and Bax immunostaining using rabbit polyclonal anti-Bcl-X L and anti-Bax antibodies (Santa Cruz Biotechnology, Santa Cruz, CA; diluted 1:100 in NGS).

Techniques: Electron Microscopy, Immunolabeling

Journal: Scientific Reports

Article Title: Evolutionary conservation of a regulative pathway of erythropoiesis in Poikilothermic vertebrates

doi: 10.1038/s41598-022-06617-6

Figure Lengend Snippet: Quantifying immunogold labelling in transmission electron microscopy and western blotting. ( a ) In torpedo erythrocytes a significant decrease of Bcl-2 and Bcl-X L and a significant increase of Bax were detected after 90 Gy irradiation (p < 0.5). ( b , c ) Quantifying immunogold labelling in young and in old loggerhead erythrocytes and in mitochondria of Torpedo marmorata Risso . A significant increase of Bax and a significant decrease of Bcl-2 and Bcl-X L were detected in old loggerhead erythrocytes, as compared to young cells (p < 0.5), in agreement with light microscopy images. ( d ) Western blotting showing apoptotic proteins involved in erythropoiesis in young and old Torpedo erythrocytes. Bcl-2 was expressed mainly in young erythrocytes. On the contrary, Bax was detected in mature red cells. After irradiation, young erythrocytes (Ir. Young) and mature erythrocytes (Ir. Old) express both Bax and Bcl-2. Moreover, cleaved caspases-3/7 were expressed at higher levels in mature erythrocytes, compared to young ones. The β-actin was used as control.

Article Snippet: The same procedure was followed for Bcl-X L and Bax immunostaining using rabbit polyclonal anti-Bcl-X L and anti-Bax antibodies (Santa Cruz Biotechnology, Santa Cruz, CA; diluted 1:100 in NGS).

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Irradiation, Light Microscopy