Figure 3.
NLRP4 is required for the recruitment of ARHGDIA to GcAV. (A) HeLa cells expressing either miR-Control (Control) or miR-NLRP4 (NLRP4 KD) were infected with GAS for 2 h. ARHGDIA and ubiquitin were immunostained. Cellular and bacterial DNA were stained with DAPI. Scale bars: 10 μm. (B) Percentages of cells with ARHGDIA-positive GAS were quantified at 2 and 4 h after infection. Data represent the mean ± SD (n > 100 cells, **P < 0.01) from 3 independent experiments. (C) Schematic representation of the ARHGDIA domain structure. (D) FLAG-NLRP4 binding to each GST-ARHGDIA-domain deletion mutant. Beads carrying the indicated purified GST fusion protein were incubated with lysates from FLAG-NLRP4-expressing HeLa cells infected with GAS. (E) HeLa cells expressing mCherry-LC3 and either GFP-ARHGDIA (WT), GFP-ARHGDIA-ΔN or GFP- ARHGDIA-ΔC were infected with GAS for 2 h. Scale bars: 10 μm. (F) ARHGDIA-positive GcAV quantification. Data represent the mean ± SD (n > 50 GcAVs, **P < 0.01) from 3 independent experiments.