The objectives of this study were (1) to investigate the distribution of large (≥10 mm) follicle numbers during the estrous cycle and (2) to compare the timing of the estrus expression period after the ovarian examination between cows with one large follicle (1F) and two or more large follicles (2F) with functional corpus luteum (CL) at the ovarian examination in lactating Holstein dairy cows. In experiment 1, we performed 393 ovarian examinations by ultrasonography, addressed the existence of CL (≥20 mm) and large follicle numbers, and classified cows into 1F (n = 229) and 2F (n = 164) groups. The 1F appearance rates were beyond 75% each day during 3 to 12 d after estrus. However, 2F appearance rates were beyond 75% each day during 15 to 24 d after estrus. In experiment 2, we performed 302 ovarian examinations by ultrasonography and classified cows into the 1F (n = 168) and 2F (n = 134) groups. Estrus detection was performed for 24 d after the ovarian examination in each cow. In the 2F group, 75% of estrus occurred within 9 d of the ovarian examination. However, 75% of estrus occurred 10 d after the ovarian examination in 1F. Days from the ovarian examination to estrus were significantly shorter in the 2F (6.0 d; median, 7.2 ± 4.0 d; mean ± SD) than in the 1F (13 d, 12.4 ± 4.3 d) group. In conclusion, focusing on ≥10 mm follicle numbers with CL could be useful for predicting the estrus expression period.