Figure 2 | Scientific Reports

Figure 2

From: Micropeptide AF127577.4-ORF hidden in a lncRNA diminishes glioblastoma cell proliferation via the modulation of ERK2/METTL3 interaction

Figure 2

The AF127577.4 ORF encodes a micropeptide AF127577.4-ORF. (A) Expression confirmation of AC090114.2-ORF, AL158166.2-ORF and AF127577.4-ORF in 293 T cells by anti-Flag antibody. (B) Viability of LN229 cells transfected as indicated was scored by CCK8 method. (C) Schematic of the AF127577.4 ORF and its coding peptide sequence. (D) Schematic of construction of AF127577.4-ORF and AF127577.4-ORFmut plasmids. (E) Detection of the AF127577.4-ORF-Flag fusion peptide in GBM cells after introduction by AF127577.4-ORF or AF127577.4-ORFmut plasmid by IB immunoblotted with anti-Flag antibody. (F) Detection of the endogenous AF127577.4-ORF in LN229 and U251 cells by IF microscopy using anti-AF127577.4-ORF antibody. Scale bars: 100 μm. (G, H) Representative IF images revealing the expression of AF127577.4-ORF in GBM cells transfected as indicated with anti-AF127577.4-ORF antibody. Scale bars: 100 μm. (I) RT-qPCR was employed to determine the knockdown efficiency of si-lncRNA AF127577.4 in LN229 cells. (J) WB assay was employed to measure the level of AF127577.4-ORF in LN229 cells transfected with vector, AF127577.4-ORF, si-NC, si-lncRNA AF127577.4, lenti-CRISPER v2 vector, and AF127577.4-ORF-CRISPER using its specific antibody.

Back to article page