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. 2021 Mar 2;12(2):465–487. doi: 10.1016/j.jcmgh.2021.02.010

Figure 9.

Figure 9

NONO mediates retention of IFNGR1 mRNA in paraspeckle of HCC cells. (A) RIP analysis of S1-tagged NEAT1_2 interacting with IFNGR1 mRNA in S1-tagged HCCML3 cells treated with proteinase K or not. (B) RIP analysis of S1-tagged NEAT1_2 interacting with IFNGR1 mRNA in S1-tagged HCCML3 cells transfected with si-NONO or si-NC. (C) RIP analysis of interacting regions of NEAT1_2 with NONO in HCCML3 cells. (D) qRT-PCR analysis of region 13-15 of NEAT1_2 expression in region 13-15 of NEAT1_2-KO HCCML3 cells. (E) Proliferation of WT or region 13-15 of NEAT1_2-KO HCCLM3 or SKhep1 cells was detected by MTT assay. (F) Apoptosis of WT or region 13-15 of NEAT1_2-KO HCCLM3 or SKhep1 cells was detected by flow cytometry. (G) RIP analysis of NEAT1_2 interacting with IFNGR1 mRNA in region 13-15 of NEAT1_2-KO HCCML3 cells using streptavidin. (H) Diagram of NONO domains (left). RIP analysis of interacting domains of NONO with IFNGR1 mRNA using anti-Flag antibody (right). (I) RIP analysis of NONO interacting with IFNGR1 mRNA in region 13-15 of NEAT1_2-KO HCCML3 cells using anti-NONO antibody. (J) Western blotting analysis of phosphorylated modifications of immunoprecipitated NONO using anti-NONO antibody in HCCML3 cells transfected with si-NONO or si-NC. (K) Western blotting analysis of phosphorylated modifications of immunoprecipitated Flag-NONO mutant using anti-Flag antibody in HCCML3 cells transfected with pcmv-Flag-NONO mutant vector and si-NONO or si-NC. (L) RIP analysis of NONO interacting with IFNGR1 mRNA in HCCML3 cells transfected with pcmv-Flag-NONO (WT) or NONO mutant using anti-Flag antibody. Data are represented as means ± SD (n = 3; ∗P < .05, ∗∗P < .01).