Light-emitting diodes (LEDs) are currently undergoing rapid development as plant growth light sources in a plant factory with artificial lighting (PFAL). However, little is known about the effects of supplementary light and pulsed LEDs on plant growth, bioactive compound productions, and energy efficiency in lettuce. In this study, we aimed to determine the effects of supplementary white LEDs (study I) and pulsed LEDs (study II) on red leaf lettuce (Lactuca sativa L. ‘Sunmang’). In study I, six LED sources were used to determine the effects of supplementary white LEDs (RGB 7:1:1, 7:1:2, RWB 7:1:2, 7:2:1, 8:1:1, 8:2:0 [based on chip number] on lettuce). Fluorescent lamps were used as the control. In study II, pulsed RWB 7:2:1 LED treatments (30, 10, 1 kHz with a 50 or 75% duty ratio) were applied to lettuce. In study I, the application of red and blue fractions improved plant growth characteristics and the accumulation of antioxidant phenolic compounds, respectively. In addition, the application of green light increased plant growth, including the fresh and dry weights of shoots and roots, as well as leaf area. However, the substitution of green LEDs with white LEDs induced approximately 3.4-times higher light and energy use efficiency. In study II, the growth characteristics and photosynthesis of lettuce were affected by various combinations of duty ratio and frequency. In particular, biomass under a 1 kHz 75% duty ratio of pulsed LEDs was not significantly different from that of the control (continuous LEDs). Moreover, no significant difference in leaf photosynthetic rate was observed between any pulsed LED treatment utilizing a 75% duty ratio versus continuous LEDs. However, some pulsed LED treatments may potentially improve light and energy use efficiency compared to continuous LEDs. These results suggest that the fraction of red, blue, and green wavelengths of LEDs is an important factor for plant growth and the biosynthesis of bioactive compounds in lettuce and that supplementary white LEDs (based on a combination of red and blue LEDs) might be more suitable as a commercial lighting source than green LEDs. In addition, the use of suitable pulses of LEDs might save energy while inducing plant growth similar to that under continuous LEDs. Our findings provide important basic information for designing optimal light sources for use in a PFAL.