Fig 1: Effect of aSyn on melanosome transfer to keratinocytes. (A) FACS analysis of the uptake of fluorescent melanosomes and changes in the granularity of keratinocytes treated with isolated and stained melanosomes. (B) Bright-field image and immunofluorescence staining of co-cultured keratinocytes and melanocytes treated with siCTR or siSNCA using TYRP1 (green) and anti-keratin14 (KRT14, red) antibodies. DAPI (blue) was used to stain the nucleus. Black and yellow arrows show melanosomes absorbed by keratinocytes, scale bars represent 20 µm. (C) Analysis of the intracellular melanin content of keratinocytes that were co-cultured with si-treated NHEMs and differentially trypsinized (n = 3). The box plots depict the mean ± SEM. Paired t-test: *: p < 0.05.
Fig 2: Full thickness conjunctival models (FTConM) displayed in vivo marker expression. Immunofluorescence staining of CK1, CK13, CK14, CK19, Vimentin, and E-Cadherin performed on FTConM after 10 days, 15 days, and 20 days of culture. Blue = DAPI, green = respective Cytokeratin/Vimentin, red = E-Cadherin. Scale bar = 100 µm.
Fig 3: Conjunctival cells maturate at the air–liquid interface and form a multi-layered epithelium. Evaluation of different media on reconstructed human conjunctival epithelium (rhConE). (A) Histological analysis via hematoxylin & eosin (HE) stainings of rhConE cultured in Pro3, E3, or A3 medium after 11 days, 15 days, and 20 days. Scale bar = 100 µm. (B) IF of CK1, CK13, CK14, CK19 performed on rhConE cultured in Pro3, E3, or A3 after 15 days of culture. Blue = DAPI, green = respective cytokeratin. Scale bar = 100 µm. (C) MTT-assay of rhConE cultured in E3 or A3 medium normalized to models cultured in Pro3 (n = 3). (D) Measurement of transepithelial electrical resistance (TEER) 1000 Hz of rhConE cultured in Pro3, E3, or A3 from 6 days until 20 days of culture (n = 3). *p < 0.05, ***p < 0.001.
Fig 4: Medium supplementation on rhConE does not induce goblet cell differentiation. Media test of Pro3 vs. Pro10 on rhConE. (A) Histological analysis of in Pro3 or Pro10 cultured rhConE via H&E (left two columns) and AB/P (right two columns) after 10 days, 15 days, and 20 days of culture. (B) Immunofluorescence of CK1, CK13, CK14, and CK19 performed on rhConE cultured in Pro10 after 15 days. Blue = DAPI, green = respective cytokeratin. Scale bar = 100 µm. (C) MTT-assay of Pro3 vs. Pro10 cultured rhConE measured via optical density of 570 nm extinction (n = 3). (D) TEER1000 Hz measurement of rhConE cultured in Pro3 or in Pro10 medium over the culture time from 6 days until 20 days (n = 3).
Supplier Page from MilliporeSigma for Anti-KRT14 antibody produced in rabbit