Fig. 2.

CRISPR/Cas9 mediated knock out of RNase2 gene in THP1-derived macrophages. A Scheme of the mutation of RNase2 caused by sgRNA1, the sequence was validated by Sanger sequencing; replacement is indicated: red labelled sequence in wild type was replaced by the green labelled sequence, resulting in the coding frame change and stop codon insertion; B western blot assay was applied to detect RNase2 protein; C secreted RNase2 in supernatant was measured by ELISA, the RPMI + 10%FBS complete culture medium was used as a negative control, the supernatant was concentrated 50 × ; D ribonuclease activity staining assay was used to confirm the removal of catalytic function. Cells were collected and resuspended in water and sonicated, cell lysates were loaded in each well at the indicated quantity