ZFIN ID: ZDB-FIG-200814-43
Tao et al., 2020 - LIN28B regulates transcription and potentiates MYCN-induced neuroblastoma through binding to ZNF143 at target gene promotors. Proceedings of the National Academy of Sciences of the United States of America   117(28):16516-16526 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Term:
Stage: Adult
PHENOTYPE:
Fish:
Observed In:
Stage: Adult

Fig. 6 Expression of WT or mutant LIN28B reverses the cell invasion and migration defects induced by GSK3B inhibition. (A) qRT-PCR to detect LIN28B, GSK3B, L1CAM, and ETV1 expression in BE2C-TET cells that were either untreated or treated with 50-ng/mL doxycycline for 3 d. Values were normalized to EEF1A1, ACTB, and B2M and represent the means ± SD of triplicate experiments. (B–D) The expression of zebrafish gsk3b (B), l1.1 (C) and etv1 (D) mRNAs in EGFP;MYCN (M), LIN28B_WT;MYCN (WT), and LIN28B_MU;MYCN (MU) tumors was analyzed by qRT-PCR. Five independent tumors from 6-mo-old fish were analyzed for each group. Each dot represents the expression levels of the indicated gene normalized to EEF1A1. Horizontal bars indicate means ± SD. (E) The LIN28B- and ZNF143-binding sequences within the GSK3B promoter region were cloned upstream of the firefly luciferase reporter gene and transfected into BE2C or BE2C-TET cells expressing Flag-tagged LIN28B_WT (WT) or LIN28B_MU (MU) that were untreated or treated with doxycycline. A reporter construct encoding renilla luciferase under control of the cytomegalovirus promoter was cotransfected to normalize firefly luciferase values. Luciferase activities were measured 48-h posttransfection and expressed as a ratio of the cells treated with doxycycline to untreated controls. Values represent means ± SEM of triplicate experiments. (F and G) Transwell migration and invasion assay of the BE2C-TET cells untreated or treated with 50-ng/mL doxycycline or 25-μM GSK3B inhibitor SB415286. Cells were stained with 0.1% crystal violet (F), and the number of migrated cells through the membrane per field was plotted as means ± SD of triplicate experiments (G). (Scale bar, 100 µm.) All of the statistical analyses were performed using the two-tailed unpaired t test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
etv1 zdf16Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
zdf16Tg; zdf39Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
zdf16Tg; zdf40Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
gsk3ba zdf16Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
zdf16Tg; zdf39Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
zdf16Tg; zdf40Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
l1camb zdf16Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
zdf16Tg; zdf39Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
zdf16Tg; zdf40Tg standard conditions Adult interrenal gland neuroblastoma RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
zdf16Tg; zdf39Tg standard conditions Adult interrenal gland neuroblastoma etv1 expression increased amount, abnormal
Adult interrenal gland neuroblastoma gsk3ba expression increased amount, abnormal
Adult interrenal gland neuroblastoma l1camb expression increased amount, abnormal
zdf16Tg; zdf40Tg standard conditions Adult interrenal gland neuroblastoma etv1 expression increased amount, abnormal
Adult interrenal gland neuroblastoma gsk3ba expression increased amount, abnormal
Adult interrenal gland neuroblastoma l1camb expression increased amount, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA