Negative values included when assessing air quality In computing average pollutant concentrations, EPA includes recorded values that are below zero. EPA advised that this is consistent with NEPM AAQ procedures. Logically, however, the lowest possible value for air pollutant concentrations is zero. Either it is present, even if in very small amounts, or it is not. Negative values are an artefact of the measurement and recording process. Leaving negative values in the data introduces a negative bias, which potentially under represents actual concentrations of pollutants. We noted a considerable number of negative values recorded. For example, in 2016, negative values comprised 5.3 per cent of recorded hourly PM2.5 values, and 1.3 per cent of hourly PM10 values. When we excluded negative values from the calculation of one‐day averages, there were five more exceedance days for PM2.5 and one more for PM10 during 2016.

It is clear from the use of ES2 and RMG-II cell lines that the Atlas Antibodies ARID1A antibody is specific for ARID1A in both Western blots and formalin-fixed paraffin embedded preparations of human origin and, coupled with the literature evidence, that it is validated in human tissue.

A No Primary antibody control (NPA) showed no staining in the epithelial or nuclear compartment (Figure 3B; Dataset e).

There was no cytoplasmic or extracellular stromal background staining present and the antibody titrated successfully losing the intensity of staining, as expected

Control slides, omitting the primary antibody, were negative except for the ER2 condition in the RMG-II cell pellet where a weak cytoplasmic background could be seen (Figure 2; Dataset d). Thus there was minimal background inherent in the staining procedure. It was therefore determined that the antibody showed specificity for formalin-fixed paraffin embedded tissues and could be run on murine tissue.

Using Western blot and IHC on murine wild-type and knockout tissue we have demonstrated that this antibody to ARID1A correctly stains murine tissue by immunohistochemistry.