Roti gelstain red

The genomic DNA of the ten Chroococcidiopsis sp. strains was extracted using a PowerWater DNA Isolation Kit (MO BIO Laboratories, Carlsbad, CA, United States) and quantified using the NanoDrop Lite Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, United States). Amplification of apcE2 was performed by using 12 ng of genomic DNA in 12 μl PCR reaction mixtures containing 6 μl of Phusion High-Fidelity PCR Master Mix with HF Buffer (New England Biolabs, Ipswich, MA, United States) and F-apcE2M and R-apcE2M primers as described (Antonaru et al., 2020 (link)). The resulting amplicons were loaded onto a 1.5% agarose gel containing 0.5 mg ml^–1 ethidium bromide and, after electrophoresis, visualized with a transilluminator.
For psaA2 and psaB2, PCR involved 50 ng of gDNA in 25 μl reactions, using a Q5 High-Fidelity 2X Master Mix (New England Biolabs, Ipswich, MA, United States). Primers are listed in Supplementary Table 1. Thermocycling conditions were as reported above, except for a 1 min extension time, and a gradient annealing temperature as described below. The results were visualized on a 1.5% agarose gel stained with ROTI GelStain Red (Carl Roth GmbH, Karlsruhe, Germany).

HEPES (4-(2-hydroxyethyl)–1-piperazineethanesulfonic acid), PEI 25 kDa, heparin sodium salt, Paraformaldehyde solution, FluorSave™ Reagent, Eagle’s Minimum Essential Medium (EMEM), RPMI-1640 Medium, fetal bovine serum (FBS), Penicillin-Streptomycin solution, Dulbecco’s Phosphate Buffered Saline (PBS), trypsin-EDTA solution, 200 mM L-glutamine solution, Paraformaldehyde, Tween20, agarose and Alcian Blue solution (1% in 3% acetic acid pH 2.5) were purchased from Sigma-Aldrich (Darmstadt, Germany). Lipofectamine 2000, SYBR gold dye, AF488-anti-rabbit secondary antibody, rhodamine phalloidin, 4′,6–diamidino–2-phenylindole dihydrochloride (DAPI), Alexa Fluor™ 647 NHS ester and Alexa Fluor™ 488 NHS ester were obtained from Life technologies (Carlsbad, California, USA). Transwell® polyester membrane cell culture inserts (0.4 μm pore size) were purchased from Corning (New York, USA). PneumaCult ALI differentiation medium, hydrocortisone and heparin were purchased from Stemcell Technologies (Vancouver, Canada). Alveofact was purchased from Lyomark Pharma (Oberhaching, Germany). ROTI®GelStain Red and bovine serum albumin were purchased from Carl Roth GmbH (Karlsruhe, Germany). Dicer substrate double-stranded siRNA (DsiRNA) targeting human GAPDH, non-specific DsiRNA and amine-modified siRNA were purchased from integrated DNA Technologies (Leuven, Belgium).

All oligonucleotides were purchased from Biomers. The sequences are listed in Table 1, with the mismatches on the RCA template marked in bold. T4 DNA ligase and 10X ligation buffer were purchased from Thermo Fisher. Φ29 polymerase and 10X polymerase buffer were purchased from Biosearch Technologies. Tris and boric acid were purchased from Thermo Fisher. EDTA disodium dihydrate was purchased from Sigma. ROTI GelStain Red was purchased from Carl Roth.