Differentiated instruction (DI) is widely promoted for addressing students’ diverse readiness, interests, and learning profiles in mathematics classrooms; however, evidence of its effectiveness remains dominated by face-to-face settings. This study examined the effect of differentiated instruction on senior secondary school students’ mathematics achievement, with gender as a moderating variable, and considered its implications for distance and digital mathematics learning. A pretest-posttest non-equivalent control group quasi-experimental design involved two intact classes from two public secondary schools in the Ijebu Education Zone, Ogun State, Nigeria. The experimental group (n = 46) received differentiated instruction, while the control group (n = 59) received conventional lecture-based instruction. Data were collected using a 50-item Mathematics Achievement Test with a KR-20 reliability coefficient of 0.84 and analyzed using descriptive statistics and ANCOVA at the .05 significance level. Results showed a significant treatment effect on mathematics achievement (F(1, 100) = 13.653, p < .001, partial η² = .120), while gender and the treatment-by-gender interaction were not significant. Supplementary document analysis indicated that readiness-, interest-, and product-based differentiation can be adapted to digital formats, whereas teacher-observed flexible grouping requires structured, data-informed alternatives. These findings demonstrate the effectiveness and gender inclusivity of DI while providing practical didactical insights for designing adaptive, responsive, and digitally mediated mathematics learning environments in distance education contexts.