Fig. 5

H89 attenuates NaCl-induced hyperosmotic stress effects on hiPSC-derived RGCs. a Immunofluorescence staining of surface and intracellular TRPV1 in hiPSC-derived RGCs treated with 50 mM NaCl with or without 5 μM H89 for 24 h. b The signal intensity in a was quantified using ImageJ and is shown in the bar graph with standard deviation. c Western blot profile of cellular responses in hiPSC-derived RGCs treated with 50 mM NaCl solution with or without 5 μM H89 pretreatment. d Immunofluorescence staining of cleaved caspase 3 performed to evaluate cell apoptosis. e The percentage of cleaved caspase 3-positive cells was quantified and is demonstrated in the bar graph. f LDH released from damaged cell was measured to evaluate the cytotoxicity; the results are demonstrated in the bar graph. g Immunofluorescent staining of BDNF expressed after the indicated treatments. h Schematic illustrating the interplay between TRPV1, PKA, CREB, and restored BDNF secretion under hyperosmotic-stress-induced damage. Asterisk (*) indicates a significant difference (P < .05)