How do I answer this question? 1c. Write a brief analysis and discussion of all

Question

How do I answer this question?

1c. Write a brief analysis and discussion of all aspects of Figure 1. Be sure to note each component (there are many!) of this figure and use information from the results to assist you in the description of these results.

Untreated DMSO Marimastat 0C 800 800 600 400 200 Uninfected Mtb Untreated DMSO Marimastat Mtb-++ Mstat 0 50 200 0 50 200 Untreated DMSO Marimastat FIGURE 1| Effect of marimastat on early granuloma formation in Mtb-infected lung tissue models. (A) Tissue models (nuclei stained with DAPI, blue) were left untreated or incubated with DMSO or marimastat before implantation with PKH26-labeled monocytes (red) and macrophages, uninfected or infected with GFP-expressing (green) H37Rv. The tissues were fixed at day 7 and subjected to DAPI-staining followed by confocal microscopy. Images show merged confocal images of randomly selected fields (uninfected models) or bacteria-harboring fields selected in the green channel (H37Rv-GFP-infected models) with treatments as indicated. (B) Z-stacks were obtained in the red channel and the volume of the cells/clusters was determined in uninfected and infected samples with or without 200 nM marmastat n = 13 C) Two different concentrations of marimastat (Mstat), 50 and 200 nM were tested n 3 (D) Regions of interest (ROIs) with (ROIbact) and without bacteria (ROlcon) were selected in the green channel and the MFI in the red channel was determined. Recruitment of monocytes/macrophages to the site of infection is expressed as ratio of MFI (ROlbact/MFI (ROlcon, n 6). Data are expressed as means +SEM. Statistical analyses were carried out using 2-way ANOVA with Sidak's multiple comparisons test for comparisons within and between groups (B), 1-way ANOVA with Tukey's multiple comparisons test within the infected group (C) and Wilcoxon signed rank test to compare MFI ratio (D). Scale bars are 100 m. ,p 0.05, ,t"p 0.0001

Explanation / Answer

This present result is of an experiment designed to understand the role of a drug Marismat on MtB infection (tuberculosis). For this experiment, they took lung-tissues created two sets- infected (experimental set) and non-infected (control set). These were then treated with the drug Marismat (200nM). Untreated tissues and DMSO treated tissues acted as control. DMSO control is used because the drugs are mostly dissolved in DMSO, so it helps to rule out any toxicity contributed by DMSO.

Now, in MtB infection, the host macrophages are affected, therefore these were observed for in the confocal imaging experiment using red and green signals. The green signal arose from the GFP present in the H37Rv strain of MtB that affected macrophages.

The confocal images were used to estimate the cell volumes using pixel intensities and as expected those tissues which were treated by the drug showed lesser number of cell volume (less green intensity) due to killing of the MtBH37Rvby the drug. With increase the drug concentration form 50nm to 200nm the cell volumes further decreased. The mean fluorescence intensity (MFI) ratio of the red channels in the non-infected and infected regions also showed decreased value thus indicating MtB killing by the drug. The controls on the other hand, did not show any killing in all cases tested. All the data were found to be statistically significant.

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