Nanoscope analysis software 1

The AFM measurements were carried out using a Bioscope atomic force microscope (Bioscope resolve™ BioAFM, Bruker), coupled with an optical microscope (Leica). Fresh-frozen mouse skin was sectioned at 10 μm thickness and sections were attached to superfrost™ microscope slides. Prior to measurements, the sections were washed with phosphate buffered saline (PBS) three times to remove residual optical cutting temperature compound (OCT). 3 sebaceous glands per skin section were chosen at random, and for each gland 3 areas of 5 x 5 μm were selected. On average 30 force-curves were included for each area. The results presented are based on approximately 1000 individual measurements pooled from 4 animals per group (WT and KO). A spherical nitride tip (5μm) on a nitride lever (SAA-SPH-5UM, Bruker) was used. The Poisson’s ratio was 0.5mm, and the minimum force fit boundary and maximum force fit boundary were 30% and 70% respectively. Trigger force was 10nN and the ramp size was 10nm. The Young’s modulus was calculated by fitting the force-curves with modulus fit model Hertzian (spherical) using the Nanoscope analysis software 1.8 (Bruker).

Molecular shapes of LBPs were acquired by AFM. LBPs (1 mg/mL) were first dissolved in ultrapure water (Milli-Q level) and then incubated at 80 °C for 2 h with continuous stirring. Sodium dodecyl sulfate (SDS) was selected as a hydrogen bond cleavage agent to disperse the entangled polymer chain. After cooling to room temperature, the solution was diluted with SDS until the solution contained an equal mass of LBPs and SDS at a concentration of 1 μg/mL. The diluted solution was then stirred for 24 h at room temperature and filtered through a 0.22 μm filter. A 10 μL aliquot of sample solution was dropped onto freshly cleaved mica substrate followed by air drying prior to observation. Afterwards, the topographies of LBP fractions were obtained with an AFM (XE-70, Park Scientific Instruments, Suwon, Korea) with tapping mode in air at room temperature (humidity: 50–60%). Nanoscope Analysis software 1.8 (Bruker, Berlin, Germany) was used for image manipulation.

The microscopy analysis at each sequential step such as the pre-PANI shield, the catalyst layer at the pre-PANI, post-PANI shield and the final SCBM (including elemental mapping) was performed using field-emission scanning electron microscope (FE-SEM, GeminiSEM 500, ZEISS, Germany) with energy dispersive spectroscopy detector (EDS, Silicon Drift Detector (SDD)-X-MaxN, OXFORD). The surface morphology at pre- and post-PANI shields and the catalyst layer was also investigated using an atomic force microscope in tapping mode (AFM, DI MultiMode V SPM, Veeco Instruments Inc. UK). For clearer interpretation, the resulting topographic height data was processed using NanoScope Analysis software 1.8 (Bruker Corporation, USA).