Fig 1: Cell-death patterns of C6 cells treated with GNR-ANGI-2 under laser or sham exposure. (A) Cell cytotoxicity pattern of C6 cells preincubated with caspase (Z-VAD) inhibitor or RIPK1 (Nec-1) inhibitor for 4 h and irradiated with laser (12 min); cell viability was evaluated using the MTT assay; cells treated with media were used as control. The bar graph represents the percentages of cell viability respective to the treatment conditions. Error bars represent the standard deviations of 3 experiments (n = 3, * p < 0.05). (B) C6 cells incubated with medium (control), laser (control+ NIR), GNR-ANGI-2 (GA), and GR-ANGI-2 (GA+NIR) were treated with 5 µg/mL AO for 30 min; then, images were recorded under fluorescence microscopy. Red fluorescence (white arrows) indicates acid vesicular organelles stained with acridine orange within the cells. (C) LC3-II processing in C6 cells treated with GNR-ANGI-2 under laser or sham exposure, and control and control treated with laser incubated for 24 h using Western blotting. (D) Changes in protein expression in C6 cells treated with GNR-ANGI-2 under sham or laser exposure. Representative Western blot results show the expression of p53, caspase-3, caspase-7, and Bcl-2 proteins. ImageJ was used to analyze the fold change in protein expression.