Inhibition of HIV-1 infection by MELK depletion with three different MELK-targeting shRNAs, 293-MELK-KD-1, 293-MELK-KD-2 and 293-MELK-KD-3, was dependent on its level of expression in the single-round infection of HEK293 cells with VSV-G pseudotyped HIV-1, reducing the likelihood of an off-target effect of the shRNA (S3A–S3C Fig). Here, MELK is linked to HIV-1 infection.