Panel A shows kymographs of Rab5a in astrocytes treated with DMSO, BAPTA-AM, and 2-APB for 2 hours. The images are labeled before and after 2 hours, with kymographs for both the process and soma regions. Panel B shows similar kymographs for LAMP1 under the same treatments. Panel C displays the resting condition with a full image field, Ca2+ positive amplitude, and kymographs from the region of interest (ROI) outlined in white. Panel D shows the global illumination condition with a full image field, Ca2+ positive amplitude, and kymographs from the ROI, highlighting Ca2+ positive-induced motility arrest and reversible recovery of LAMP1 motility.
Ca 2+ chelation suppresses organelle motility. (A and B) Kymographs of Rab5a (A) and LAMP1 (B) in astrocytes treated with 100 µM BAPTA-AM or 100 µM 2-APB for 2 hr. DMSO as a control. Scale bars: 10 μm (whole cell view); 5 μm (kymographs). (C and D) For each condition, panels show the full image field with the ROI outlined in a white box (left), the corresponding Ca2+ amplitude (middle), and kymographs from this ROI (right). Acquired at 0.5-s intervals using the Zeiss Airyscan Fast mode. Scale bars: 10 μm (whole cell view); 10 μm (kymographs). (C) Resting condition. Kymographs show basal LAMP1 motility over a 4-min recording window. (D) Global illumination condition. Kymographs show LAMP1 motility over an 8-min recording window following Global illumination. The black dashed box marks Ca2+-induced motility arrest, and the blue dashed box highlights reversible recovery of LAMP1 motility.