Fig 2: Overexpression of P2X7R in PDLSCs (adenovirus transduction).a P2X7R mRNA expression in PDLSCs (assessed by qRT-PCR) in response to transduction with Ad-control or Ad-P2X7 at 10 pfu/cell (MOI = 10) or 20 pfu/cell (MOI = 20); cells without transduction were used as the control group. b P2X7R protein expression in PDLSCs (assessed by western blotting) in response to transduction with Ad-control or Ad-P2X7 at 10 pfu/cell (MOI = 10) or 20 pfu/cell (MOI = 20); cells without transduction were used as the control group. c Proliferative activity of Ad-P2X7-transfected PDLSCs (MOI = 20) and Ad-control-transfected PDLSCs assessed by the CCK-8 assay. d Representative images (confocal micrograph) of PDLSCs transfected with Ad-P2X7 or Ad-control (MOI = 20), which carries the green fluorescence protein (GFP) gene and P2X7R (immunofluorescence staining with a P2X7R antibody, red) (scale bar = 20 µm). Data are presented as the mean ± S.D. for n = 3; ns indicates no significant difference between the indicated columns; *p < 0.05, **p < 0.01, and ***p < 0.001 indicate significant differences between the indicated columns

Fig 3: Schematic displaying our proposed model for ATP release. Our proposed model suggests that ivermectin activates P2RX4 and P2RX7 leading to the release of ATP and the more ATP that accumulates extracellular can promote cell death especially in the presence of paclitaxel. In addition, the breakdown of ATP can be prevented in the presence of E-NTPDase inhibitors POM-1 or PSB 069. However, the release of ATP can be prevented in the presence of P2RX4 inhibitors 5-BDBD or Iso-PPADS or P2RX7 inhibitors A438079 or Iso-PPADS.

Fig 4: P2X7R expression in PDLSCs under normal/osteogenic conditions with or without the addition of inflammatory factors.a Quantitative analysis of P2X7R mRNA expression in PDLSCs in normal a-MEM culture medium, inflammatory culture medium (medium supplemented with TNF-a and IL-1ß), osteo-inductive culture medium, and inflammatory osteo-inductive culture medium (osteo-inductive culture medium supplemented with TNF-a and IL-1ß). b Western blotting of P2X7R in PDLSCs in normal a-MEM culture medium, inflammatory culture medium (medium supplemented with TNF-a and IL-1ß), osteo-inductive culture medium, and inflammatory osteo-inductive culture medium (osteo-inductive culture medium supplemented with TNF-a and IL-1ß). Data are presented as the mean ± S.D. for n from 4 to 6; ns indicates no significant difference between the indicated columns; *p < 0.05, **p < 0.01, and ***p < 0.001 indicate significant differences between the indicated columns

Fig 5: Local inhibition of P2X7R in Vc reduces HS-evoked OOemg activity during DED. The selective P2X7R antagonist (A804598) was microinjected (10 µM, 0.2 µL) into the Vc of sham and 14 days DED rats 30 min prior to the test stimulus (ocular instillation of 2.5 M NaCl). (A) Males, (B) OvX females, (C) OvXE. Gray bars = sham; red bars = 14-day DED. *p < 0.01, p < 0.001 versus 0.15 M NaCl; b = p< 0.01, c = p< 0.001 versus pre-drug. N = 10 rats per group.