Fig. 5

Reduced NETosis in SNHG16- or miR-146a-silenced HL-60 cells. a Map of pAll-EF1a-CasRx (16,014 bp in length) with an EGFP insertion, a Cas13d domain, a 2.7 k bp stuffer and a 30 bp DR with an appended G. b Upper, flow cytometric graphs of sorted GFP-positive CasRx-transduced cell portion for qRT-PCR analyses including CasRx-01 to 09- and CasRx-NC-transfected 293 T cells. Lower, SNHG16 levels for silencing efficacy in CasRx-01 to 09- and CasRx-NC-transfected 293 T cells. c SNHG16 and miR-146a levels in CasRx-01 to 05- and CasRx-NC-transfected 293 T cells. d SNHG16 and miR-146a levels in CasRx-02-transfected HL-60 cells. e Quantification of NETs formation percentages (left), CitH3 levels (middle), and PAD4 levels (right) in SNHG16-silenced dHL-60 cells under PMA stimulation. f Quantification of NETs formation percentages (left), CitH3 levels (middle), and PAD4 levels (right) in PMA-stimulated CasRx-02-transfected dHL-60 cells in which miR-146a was silenced by creating sh-miR-146a-transfected stable transfectants. Values are mean ± SEM. All results in this figure were representative of at least 2 independent experiments with similar findings. * p < 0.05, ** p < 0.01, *** p < 0.001. DR: direct repeat, HA: human influenza hemagglutinin amino acids 98–106. NLS: nuclear localization sequence