Fig. 4
Localization and expression of PINT87aa in cells and human tissues. a IF microscopy images of the cellular localization of the PINT87aa-RFP-fusion protein in 456 and 4121 BTIC cells showing stable overexpression. Scale bars, 20 μM. b The expression of circPINTexon2 and PINT87aa was detected in human brain, breast, liver, kidney, stomach, intestine, thyroid, and pancreas tissues. c Upper panel, the expression of circPINTexon2 and PINT87aa was detected in hNSC, 293T, Hs683, SW1783, 4121, 456, 387, and H2S cells. Lower panel, circPINTexon2 and PINT87aa levels were determined in normal brain and gliomas with different WHO grades. d Establishment of linear vector-transduced stable PINT87aa-GFP 456 and 4121 cells and artificial circPINTexon2-transduced stable PINT87aa 456 and 4121 cells. Data are presented as mean ± s.e.m. from three independent experiments. **P < 0.01; ns, P > 0.05, determined by two-tailed Student’s t-tests. e Establishment of PINT87aa K.O. SW1783 and Hs683 cells using CRISPR/Cas9 technology. Schematic illustration showing that the genomic regions of the LINC-PINT exon2 contained the 87-aa ORF. gRNAs designed to target the PINT87aa ORF are shown. Genomic K.O. effects were confirmed via Sanger sequencing, as shown in Supplementary Fig. 7. Western blotting revealed the effects of PINT87aa K.O. in SW1783 and Hs683 cells
