Fig 2: Knockdown of ubiquitin-associated protein 2-like (UBAP2L) by lentivirus-mediated short hairpin RNA (shRNA) in glioma cells. (A) Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis of UBAP2L expression in U251, U87MG, A172, U373 and U-118MG glioma cells. (B) Determination of infection efficiency in human glioma cells. Representative images of U251 and U373 cells after 4 days of lentivirus infection are shown. (C and D) Expression analysis of UBAP2L mRNA and (E and F) protein levels in U251 and U373 cells by RT-qPCR and western blot analysis. The β-actin gene and the GAPDH protein were the internal controls for RT-qPCR and western blot analysis, respectively. Con, cells without infection; Lv-shCon, cells infected with the Lv-shCon; Lv-shUBAP2L, cells infected with the Lv-shUBAP2L. Scale bar, 100 µm.

Fig 3: Determination of silencing efficiency of lentivirus-mediated RNA interference against UBAP2L in breast cancer cells. Knockdown of UBAP2L by shRNA was confirmed by a quantitative real-time PCR and b Western blotting analysis. Data represent the mean ± SD of triplicate samples. GAPDH was used as internal control. **P < 0.01, ***P < 0.001 vs. negative control (NC)

Fig 4: Haploinsufficiency of Ubap2l in mouse leads to behavioral and cognitive impairments.(A) Targeting strategy of Ubap2l conventional KO mice. (B) The percentage of mice with different genotypes [WT, heterozygous (HET), and homozygous (HOM)] at E15.5, E18.5, and P1. (C) The three-chamber test for the WT and HET mice. The time spent with empty (E), stranger 1 (S1), and stranger 2 (S2) of the WT and HET mice was recorded and compared. The preference indexes [(S1 − E)/(S1 + E) and (S2 − S1)/(S1 + S2)] were calculated and compared (n = 24 for WT and n = 22 for HET). (D) The Y-maze test for the WT and HET mice. The percentages of spontaneous alternation behavior of the WT and HET mice were calculated and compared (n = 11 for WT and n = 10 for HET). (E) The open-field test for the WT and HET mice. Time spent in the center zone and traveled distance of the WT and HET mice were recorded and compared, respectively (n = 31 for WT and n = 27 for HET). (F) The elevated-plus maze test for the WT and HET mice. Time spent and distance traveled in the open arms and closed arms of the WT and HET mice were recorded and compared, respectively (n = 23 for WT and n = 21 for HET). *P < 0.05 and **P < 0.01.

Fig 5: Silencing of UBAP2L induced G2/M phase arrest in breast cancer cells. a The percentage of cell cycle distribution was analyzed using flow cytometry. Statistical analysis showed that more cells were accumulated at G2/M phase in shUBAP2L-2 group as compared with the control group in ZR-75-30 and T-47D cells. b Differences in protein expression level of Cyclin B1, CDK1 and P21 between shUBAP2L-2 and NC or blank groups were determined by Western blot in ZR-75-30 and T-47D cells. Data represent the mean ± SD of triplicate samples. *P < 0.05, ***P < 0.01, ***P < 0.001 vs. negative control (NC)