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The continued warming of the Arctic could release vast stores of carbon into the atmosphere from high-latitude ecosystems, especially from thawing permafrost. Increasing uptake of carbon dioxide (CO2) by vegetation during longer growing seasons may partially offset such release of carbon. However, evidence of significant net annual release of carbon from site-level observations and model simulations across tundra ecosystems has been inconclusive. To address this knowledge gap, we combined top-down observations of atmospheric CO2 concentration enhancements from aircraft and a tall tower, which integrate ecosystem exchange over large regions, with bottom-up observed CO2 fluxes from tundra environments and found that the Alaska North Slope is not a consistent net source nor net sink of CO2 to the atmosphere (ranging from -6 to +6TgCyr-1 for 2012-2017). Our analysis suggests that significant biogenic CO2 fluxes from unfrozen terrestrial soils, and likely inland waters, during the early cold season (September-December) are major factors in determining the net annual carbon balance of the North Slope, implying strong sensitivity to the rapidly warming freeze-up period. At the regional level, we find no evidence of the previously reported large late-cold-season (January-April) CO2 emissions to the atmosphere during the study period. Despite the importance of the cold-season CO2 emissions to the annual total, the interannual variability in the net CO2 flux is driven by the variability in growing season fluxes. During the growing season, the regional net CO2 flux is also highly sensitive to the distribution of tundra vegetation types throughout the North Slope. This study shows that quantification and characterization of year-round CO2 fluxes from the heterogeneous terrestrial and aquatic ecosystems in the Arctic using both site-level and atmospheric observations are important to accurately project the Earth system response to future warming.