CP26 is a monomeric minor light-harvesting complex of Photosystem II (LHCII) protein located at the interface between LHCII trimers and the PSII core in thylakoid membranes. Previous studies have proposed CP26 plays a role in non-photochemical quenching (NPQ) in addition to light harvesting. Here, we utilized biophysical and pharmacological approaches to investigate this role using single- and higher-order Arabidopsis (Arabidopsis thaliana) cp26 mutants, examining its relationship to known NPQ regulators (Photosystem II subunit S, PsbS, violaxanthin de-epoxidase, VDE, and the pH gradient across the thylakoid membrane). cp26 mutants showed significantly reduced maximum PSII quantum efficiencies (F_v/F_m) in darkness, indicating a constitutively quenched state further confirmed by fluorescence lifetime measurements. Destabilized PSII-LHCII supercomplexes observed in native gel electrophoresis and tighter PSII supercomplex packing were potential causes, with no other antenna proteins capable of rescuing this phenotype. In addition, the cp26 mutants exhibited altered NPQ capacity—modest in single mutants but substantial in double mutants—independent of PsbS and VDE. Together, these results show that CP26 is not involved in qE or qZ but may primarily play an indirect role in apparent NPQ responses via PSII-LHCII supercomplex organization.