Fig 1: PCR of gDNA sample extracted from E. intestinalis ATCC 50506 spores purified from infected HFF-1 cells post DNase I treatment.Ladder: 100 bp ladder. Neg: negative control with all used primers. Well 1: universal bacterial primers for 16S gene (357F-1100R). Well 2: universal mammal primers for rRNA gene (MF787-MR7535). Wells 3-5: E. intestinalis specific primers for rRNA (lsuF1-lsuR1, lsuF2-lsuR2, and ssuF1-ssuR1). Well 6: universal microsporidian primers for small rRNA subunit (ss530F- ss1047R).
Fig 2: Encephalitozoon intestinalis ATCC 50506 spores in HFF-1 cell culture.Culture visualized under phase-contrast inverted microscope with 10X objective and Canon EOS REBEL T3i digital camera. Each spore is about 1 µm wide to 3.5 µm in length.
Fig 3: HFF-1 cells infected with Encephalitozoon hellem ATCC 50451 at different post- infection days. E. hellem vacuoles in HFF-1 cell culture (magenta arrow) at 7 dpi (A), 27 dpi (B), and 32 dpi (harvest stage) (C).
Fig 4: Amplification of bacterial 16S, mammal and microsporidian rRNA in the E. intestinalis ATCC 50506 spore sample obtained from HFF-1 cell culture.Ladder: 100 bp ladder (FroggaBio, ON, Canada). Neg: negative control, no sample. Well 1: E. intestinalis/E. hellem lsuF1-lsuR1 primers. Well 2: E. intestinalis/E. hellem lsuF2-lsuR2 primers. Well 3: E. intestinalis/E. hellem ssuF1-ssuR1 primers. Well 4: bacterial 357F-1100R primers. Well 5: mammal MF787- MR7535 primers. Bac + : Staphylococcus salivarius as positive control for bacteria primers.
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