Fig. 4
(A) CLSM images of CT26 cells treated with RCuH for 2, 4, and 6 h at 37℃, respectively. (B) Flow cytometry profiles of CT26 cells incubated with RCuH for 2, 4, and 6 h at 37℃, respectively. (C) In vitro MTT-measured toxicities of TPZ, TCu, TCuH, and TCuH + 1064 nm laser (shown as + L in the figure) for 24 h. (D) Live/dead assay of CT26 cells incubated with PBS, TPZ, CuH, TCuH and TCuH + L (100 µg/mL) for 6 h. (E) Intracellular JC-1 assay of HeLa cells after different treatments with PBS, TPZ, CuH, TCuH and TCuH + L (100 µg/mL) for 6 h. (F) Representative Bio-TEM images of CT26 cells before and after treatment with TCuH + L. Red oval indicates location of mitochondria. (G) Schematic illustration of the possible mechanism by which TCuH promotes cuproptosis. (H) Western blot analysis of expression levels of HSP70, HSP90, FDX1, ATP7A, and GPX4 after incubation with PBS, TPZ, CuH, TCuH, or TCuH + L (100 µg/mL) for 24 h. (I) Quantitative comparison of dissolved H2S levels among CT26 cells treated with PBS, TPZ, CuH, TCuH, or TCuH + L (100 µg/mL) for 24 h. (J) Quantitative comparison of dissolved ATP levels among CT26 cells treated with PBS, TPZ, CuH, TCuH, or TCuH + L (100 µg/mL) for 24 h. (K) Quantitative comparison of dissolved GSH levels among CT26 cells treated with PBS, TPZ, CuH, TCuH, or TCuH + L (100 µg/mL) for 24 h