Fig. 4: LZD increases ER-mitochondria association.
From: ER-mitochondria association negatively affects wound healing by regulating NLRP3 activation
a Mitochondrial Superoxide levels in HaCaT cells, treated as indicated, were detected by Mitosox probe (n = 3 independent experiments; Two-way ANOVA: ****P < 0.0001). b Cytosolic mtDNA evaluation by the analysis of mt-CO1 and mt-Dloop levels, in HaCaT cells, treated as indicated (n = 3 independent experiments; Two-way ANOVA: **P = 0.0014 [uninfected/untreated vs MRSA/untreated]; **P = 0.0016 [MRSA/untreated vs MRSA/Vanco]; **P = 0.0023 [MRSA/untreated vs MRSA/LZD]; **** P < 0.0001). c Representative confocal images of ER-mitochondria association in HaCaT cells, transiently transfected with SEC61-GFP (ER marker) and mitochondrial-targeted cherry (mt-cherry, mitochondrial marker), and then treated as indicated (scale bars: 5 µm). Merged images are shown. Magnification of SEC61-GFP, mt-cherry, and merged images in insets. d Quantification of ER-mitochondria associations by Manders’ coefficients calculation in HaCaT cells, treated as indicated (n = 3 independent experiments; Two-way ANOVA: **P = 0.0024 [M1: uninfected/untreated vs MRSA/untreated]; **P = 0.0080 [M1: uninfected/untreated vs uninfected/LZD]; **P = 0.0092 [M1: uninfected/Vanco vs uninfected/LZD]; ***P = 0.0005 [M1: MRSA/untreated vs MRSA/Vanco]; ***P = 0.0005 [M1: MRSA/Vanco vs MRSA/LZD]; ***P = 0.0009 [M2: MRSA/untreated vs MRSA/Vanco]; **P = 0.0012 [M2: MRSA/Vanco vs MRSA/LZD]; ****P < 0.0001). e Schematic drawing representing principles of the BRET method. Bioluminescence is developed as a consequence of the proximity of R-Luc (mitochondria) and mVenus (ER). f Quantification of bioluminescence intensity detected by BRET assay, in HaCaT cells, treated as indicated (n = 3 independent experiments; Two-way ANOVA: *P = 0.0383; ***P = 0.0008; ****P < 0.0001).
