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. 2021 Nov 17;11:756148. doi: 10.3389/fonc.2021.756148

Figure 4.

Figure 4

Lnc-THOR-driven NSCLC cell growth is through binding to IGF2BP1. Western blotting of the nuclear IGF2BP1 protein retrieved by biotin-labeled full-length Lnc-THOR in pCan-1 NSCLC cells and A549 cells (A). qRT-PCR assay of Lnc-THOR enriched by IGF2BP1 protein in pCan-1 cells and A549 cells (B). The stable pCan-1 cells expressing the CRISPR/Cas9-IGF2BP1-KO construct (“IGF2BP1-KO” cells) were further transduced with the “sh-S1” Lnc-THOR shRNA or the Lnc-THOR-expressing construct (“Lnc-THOR-OE”), control cells were transduced with the Cas9-KO empty vector (“Cas9-C”); Expression of IGF2BP1 mRNA (C), listed proteins (D) and Lnc-THOR (E) was shown; Cells were further cultured for applied time periods, cell proliferation and migration were tested by nuclear EdU staining (F) and “Transwell” (G) assays, respectively, with results quantified. The CRISPR/Cas9-edited Lnc-THOR KO pCan-1 cells (“koTHOR”) were further infected with recombinant adenovirus encoding the human IGF2BP1 expression construct (“OE-IGF2BP1”) or empty vector (“Vec”), control cells were with the Cas9-KO empty vector (“Cas9-C”); Expression of IGF2BP1 mRNA (H), listed proteins (I) and Lnc-THOR (J) was shown; Cells were further cultured for applied time periods, cell proliferation (K) and migration (L) were tested, with results quantified. Data were presented as mean ± standard deviation (SD, n=5). *P < 0.05 vs. “Cas9” cells. “N. S.” stands for non-statistical difference. The experiments were repeated five times, with similar results obtained. Scale Bar = 100 μm (F).