Fig. 3From: Identification of target genes regulated by encystation-induced transcription factor Myb2 using knockout mutagenesis in Giardia lambliaThe glmyb2-deletion mutants JK1 and JK2 show defective glcwp1 expression and cyst formation during encystation. a Quantitative PCR (qPCR) analysis of glmyb2 gene expression during encystation. Various Giardia cells (WB, JK1, and JK2) incubated in the encystation medium for 3, 6, and 14 h were used to extract RNA samples. qPCR was performed using primers specific for glmyb2 genes, and transcript levels were normalized to those of the G. lamblia actin-related gene (glactin; GL50803_15113). Three independent replicates, each consisting of three technical replicates, were evaluated. Error bars represent the standard error of the mean. b IFAs of WB, JK1, and JK2. Giardia trophozoite, 14 h and 24 h encysting cells were stained with anti-GlCWP1 antibodies (1:200) and then incubated with Alexa Fluor 555-conugated anti-rat IgG (1:100). The slides were mounted with ProLong™ Gold Antifade Mountant with DAPI. They were then observed with an Axiovert 200 fluorescent microscope. Differential interference contrast (DIC) image shows cell morphology. Scale bars, 2 μm. c Cyst formation of WB, JK1, and JK2 strains. Scanning electron micrograms of various Giardia strains (WB, JK1, and JK2) as trophozoites and encysting cells at 14 h and 24 h post-induction to encystation. Cysts are indicated with asterisks, and pseudocyst phenotype cells are indicated with double asterisks. The bars indicate 2 μm. d Efficiency of wild-type and mutant Giardia in cyst formation. WB, JK1, and JK2 strains were incubated in encystation medium for 48 h and treated with water, and then their cyst formation was analyzed. Three independent experiments were performed and analyzed. Error bars represent the standard error of the meanBack to article page