Figure 2.

LP-A96 promotes scratch closure and induces Ca²⁺ influx in corneal epithelial cells. (A) A scratch (gap) assay was used to observe the enhancement of cell motility. The velocity of cell monolayer towards the remaining gap area (devoid of cells) was calculated as outlined in (B). (B) Gaps were imaged before the treatment and at 12 h post-treatment, and used to construct (A). White dotted lines indicate the outer edges of the monolayer adjacent to the wound gap. (C~E) Human corneal epithelial (HCE-T) cells pre-incubated with Fluo-4 AM were incubated with either LP-A96, Lacripep, or A96 (all at 1 μM). The dotted line at the 5th minute indicates when the treatment was added to the culture medium. The fluorescence intensity of Fluo-4 AM was monitored as a marker for Ca²⁺ influx (n = 27~33 cells/treatment per experiment). A representative data set from the three independent sets of the experiment is shown. (F) The area under the curve of each fluorescence intensity profile shown in (C~E) showed that only LP-A96 extensively mobilized Ca²⁺. A one-way ANOVA followed by multiple comparisons was used for statistical comparison. **** p < 0.0001, * p < 0.05, ns: non-significant. Mean ± SD.