Bp2818500

IAA (I2886) and NAA (N0640) were obtained from Sigma‐Aldrich and diluted to a 10 mM stock concentration in ultrapure ethanol (BP2818500; FisherScientific, Waltham, MA, USA). For experiments, each stock was diluted to appropriate concentrations into ½MS liquid medium without sucrose; for control solution, ultrapure ethanol was added to ½MS liquid medium without sucrose to match the highest auxin concentration used. Because light degrades auxin (Nissen & Sutter, 1990), blinded solutions were kept wrapped in foil throughout an experiment. To ensure even treatment during 20–30 min treatments, whole seedlings were cut from agar plates and treated by soaking their agar block in a 24‐well plate. For 100‐s timelapse movies, plants were treated on slides by being mounted in either control or IAA solution. Imaging began almost immediately; timelapse movies from both Regions 2 and 3 were collected within 7 min. For 20–30 min treatments, imaging concluded within 30 min. Because darkness can stimulate degradation of cytoskeletal organizing proteins (Dyachok et al., 2011) and a reorientation of actin filaments in hypocotyls (Breuer et al., 2014), plants were left under grow lights (40 μM photons m⁻² s⁻¹) while soaking during 20‐min treatments, and slides were prepared in the light. All IAA and NAA experiments were performed and analyzed double blind.

Biopsies were consistently obtained from each breast for every study patient. During Surgery #1, and prior to the conclusion of mastectomy, ≥3 sq cm biopsies of the skin and underlying breast parenchyma were obtained (Fig. 2A) at least 3 cm away from a known tumor margin to avoid compromising pathologic assessment. After Surgery #1, one suction drain was aseptically collected in the plastic surgery outpatient clinic at the time of removal and only the portion within the breast tissue was retained for future analyses (Fig. 2B). All patients matriculated, uneventfully, to Surgery #2 where biopsies from the skin (≥3 sq cm), ADM (≥3 sq cm), TE (≥30 sq cm), capsule (≥3 sq cm), and subcutaneous tissues (≥ 3 sq cm) were obtained (Fig. 2C). Immediately upon removal, specimens intended for DNA extraction and 16S rRNA sequencing were stored in 200 proof ethanol (Fisher Scientific; BP2818500), while remaining samples were immediately placed in 4% phosphate buffered solution (PBS) for bacterial culturing and immunofluorescence staining as previously described (25 (link), 93 (link)) (also see Bacterial Culture, Bacterial Identification, Immunofluorescence Sample Processing, and Microbiome Sample Processing sections below).