The tradition of miniature painting, known as Negārgari in Persian dictionaries, plays a crucial role in Iran's visual culture and has long been regarded as a foundational pillar of Persian artistic expression. As one of the oldest and most enduring forms of Iranian visual and traditional art, miniature painting has historically functioned not only as an aesthetic endeavor but also as a vehicle for intellectual, spiritual, and literary transmission. In the broader context of Persianate art, it is often revered as the "mother of arts" due to its sophisticated integration of narrative, myth, design, and symbolic meaning. This art form, throughout the dynamic and often turbulent history of Iran, evolved through two principal trajectories: the manuscript illustration tradition, which adorned illuminated books and handwritten texts, and monumental mural painting, which embellished the interior walls of significant architectural structures. Both forms reached a particularly vibrant stage of development during the Safavid period (1501–1722), especially within the framework of the Isfahan School, which emerged as a leading center for artistic innovation and synthesis in early modern Iran. This research investigates the relationship between these two major artistic expressions, miniature painting and wall painting, through a comparative case study focusing on the murals of Ālī Qāpū Palace and the illustrated manuscripts produced under the Isfahan School. The main objective is to assess the degree of mutual influence between these two visual traditions while also exploring the impact of Western artistic elements on their stylistic and conceptual formation. By adopting a descriptive-analytical approach and relying on both field observations and library-based sources, this study provides a nuanced evaluation of how the socio-political, religious, and cultural contexts of the Safavid era shaped the evolution of these visual forms. A selection of key illustrated manuscripts representative of the Isfahan School was used as the core comparative corpus. These include the Shahnameh preserved in the Chester Beatty Library (996–1006 AH), the Shahnameh of Shah Abbas from the Spencer Collection (1023 AH), a Mantiq al-Tayr manuscript attributed to the period of Shah Abbas I, a Gulistan of Sa'di dated 1024 AH, a copy of Jāmiʿ al-Tawārīkh associated with the reign of Shah Abbas II and held in Golestan Palace, and a late Shahnameh produced during the declining years of the Safavid dynasty under Shah Sultan Hussein (Metropolitan Museum). These were studied alongside selected mural panels and decorative wall paintings from Ālī Qāpū Palace, with particular attention to iconographic motifs, compositional structure, formal elements, and narrative content. The study's findings reveal both continuities and ruptures between the two media. While miniature paintings remained closely linked to literary texts and maintained a format optimized for private viewing and aristocratic patronage, mural paintings—especially those at Ālī Qāpū—moved toward a more public, performative, and often secular aesthetic. The murals of Ālī Qāpū exhibit subjects such as musical gatherings, feasts, female figures, and Europeanized interiors, indicating a distinct shift in both content and function. These images reflect the growing importance of courtly entertainment, the increasing visibility of women in visual culture, and the subtle incorporation of Western norms—likely facilitated through Armenian merchants, diplomatic exchanges, and the missionary presence in New Julfa. Stylistically, although both art forms share a foundation in Persian aesthetics, mural painting began to deviate from traditional conventions during the late Safavid period. The use of chiaroscuro (light-and-shadow modeling), linear and atmospheric perspective, naturalistic anatomy, and European dress codes all suggest the influence of Western artistic vocabularies. Such elements are more prominently seen in the single-leaf (tak-pardeh) painting format, which flourished independently of manuscript production and is considered a transitional form between classical miniature and later Qajar portraiture. The mural works of Ālī Qāpū thus show stronger affinities with this single-sheet genre than with manuscript miniatures in the strict sense. Therefore, this research proposes that while manuscript illustration and wall painting coexisted and even overlapped in the Isfahan School, they represent divergent artistic trajectories shaped by differing material, functional, and ideological contexts. The palace murals were not simply enlarged transpositions of book miniatures; rather, they constituted a distinct visual language, responsive to architectural space, changing patronage systems, and new aesthetic expectations. This distinction challenges prevailing assumptions in Iranian art history that tend to overemphasize the unity or continuity between artistic media and underscores the need for more context-sensitive approaches to the study of Persian visual culture. Ultimately, the murals of Ālī Qāpū should be interpreted not merely as decorative supplements to architecture, nor as direct extensions of manuscript traditions, but as autonomous artistic productions shaped by Safavid court culture, urban cosmopolitanism, and cross-cultural artistic exchange. In light of these findings, the study repositions Ālī Qāpū's murals within the broader narrative of Persian art history as key markers of both continuity and transformation at the crossroads of tradition and modernity.