Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Pleiotropic Roles for the Plasmodium berghei RNA Binding Protein UIS12 in Transmission and Oocyst Maturation

doi: 10.3389/fcimb.2021.624945

Figure Lengend Snippet: Down-regulation of distinct mRNAs coding for signature gamete-, ookinete- and oocyst-specific proteins in uis12(-) blood stage parasites. Shown are mean values of microarray results of total RNA isolated from mixed infected erythrocytes from uis12(-) (clone 1) - and WT- infected mice from two biological replicates. (A) Shown is a volcano-plot illustrating the fold change of the expression levels and the negative log p -values of all analyzed 2,890 P. berghei genes. The dotted black line represents a p -value of 0.05 and all transcripts with a p -value <0.05 are shown in red. Exemplary transcripts are highlighted and labeled in blue. (B) Pie charts displaying the proportions of up- (green >2), non- (grey), and down- (orange <-2) regulated transcripts among all transcripts (upper left), blood stage-specific transcripts (center), and gametocyte-specific transcripts (upper right) ( Otto et al., 2014 ). Exemplary transcripts are listed in the respective region of the chart. The number of transcripts analyzed is shown in a white circle inside the center. (C) Pie charts displaying the proportions of up- (green >2), non- (grey), and down- (orange <-2) regulated transcripts among female (left) and male (right) gametocyte-specific transcripts ( Yeoh et al., 2017 ). The number of transcripts analyzed is shown in a white circle inside the center.

Article Snippet: Scanning of the microarrays was performed with 5 µm resolution and the extended mode using a ‘High Resolution Microarray Laser Scanner’ (G2505, Agilent Technologies).

Techniques: Microarray, Isolation, Infection, Expressing, Labeling

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Pleiotropic Roles for the Plasmodium berghei RNA Binding Protein UIS12 in Transmission and Oocyst Maturation

doi: 10.3389/fcimb.2021.624945

Figure Lengend Snippet: Confirmation of down-regulation of selected mRNAs by quantitative RT-PCR. Quantitative RT-PCR on selected blood stage and gametocyte transcripts validate the results of the microarray analysis. Shown are -fold changes of steady state mRNA levels of uis12(-) clone 1 compared to wild-type mixed blood stages for selected genes ( Puf1 , P28 , MDV , DOZI , Actin II , UIS1/IK2 , SET , MSP1 , and AMA1 ). qRT-PCR data were normalized to the steady state levels of HSP70 mRNA. The microarray data represent mean values of biological replicate 1 and 2 ( ± SD).

Article Snippet: Scanning of the microarrays was performed with 5 µm resolution and the extended mode using a ‘High Resolution Microarray Laser Scanner’ (G2505, Agilent Technologies).

Techniques: Quantitative RT-PCR, Microarray

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Pleiotropic Roles for the Plasmodium berghei RNA Binding Protein UIS12 in Transmission and Oocyst Maturation

doi: 10.3389/fcimb.2021.624945

Figure Lengend Snippet: A motif is shared by up-regulated transcripts in uis12(-) parasites, and the comparison of dozi(-), cith(-) and uis12(-) target transcripts reveals possible overlapping functions. (A) Shown is a graphic display of the 10-nucleotide U-rich motif found in the ORF of transcripts that are up-regulated in absence of UIS12 in the microarray with biological replicate 1. The size of the depicted nucleotide represents its probability at the respective position. (B) Venn diagram comparing shared down-regulated transcripts of the gametocyte microarray analyses performed for dozi(-) and cith(-) with a -fold change lower than -1 ( Mair et al., 2010 ) and down-regulated transcripts in mixed blood stages of uis12(-) with a mean -fold change lower than -2.

Article Snippet: Scanning of the microarrays was performed with 5 µm resolution and the extended mode using a ‘High Resolution Microarray Laser Scanner’ (G2505, Agilent Technologies).

Techniques: Microarray