Validation of ground-based and satellite remote sensing CO2 observations involves comparisons among platforms and with in situ airborne measurements. Several factors unrelated to observational errors can lead to mismatches between measurements, and must be assessed to avoid misinterpreting actual differences in observed values as errors. Here we explore the impact of CO2 horizontal variability and differences in the spatial support of measurements. Case studies based on flights over Walnut Grove and Petaluma, California, are used to compare hypothetical airborne, TCCON, GOSAT, and OCO-2 measurements. We find that high CO2 variability can lead to differences in inferred XCO2 (1) of over 0.5 ppm between airborne and remote sensing observations, due to the spatial mismatch between spiral flight trajectories and atmospheric columns, and (2) of up to 0.3 ppm among remote sensing platforms, due to differences in the spatial support of observations. Horizontal CO2 variability must therefore be considered in intercomparisons aimed at validation of remote sensing observations.