Fig. 2.

Detection of oxidized PTP1B by dimedone-PLA. Serum-starved HepG2 cells expressing control shRNA or shRNA targeting human PTPN1 (shPTPN1), with or without stable expression of either mouse WT PTP1B or PTP1B^C215S, were stimulated with insulin (25 nM) or H₂O₂ (1 mM) for 5 min or left unstimulated. Cells were then subjected to dimedone-PLA with anti-dimedone-Cys and anti-mouse PTP1B antibodies (gray). Representative images are shown for each condition from one of two independent biological replicates. The graph shows the average number of PLA signals per cell (n = 6 images for each condition, 80–120 cells in an image), relative to control cells without stimulation (set as 1). ***P < 0.0001, ns not significant, ANOVA with Bonferroni/Dunn’s post-hoc test. Error bars represent SD. Scale bar: 50 μm