Quantitative Proteomics and Lipidomics Analysis of Endoplasmic Reticulum of Macrophage Infected with Mycobacterium tuberculosis

Figure 2

Induction in host cell cytosolic Ca²⁺ and subsequent inhibition of SERCA upon infection with H37Ra at 24 hrs. (a) (i) Representative image of western blot of ATP2A2 upon infection with H37Ra and H37Rv probed with anti-ATP2A2 primary antibody where β-actin was used as a loading control. Lysates from UI, H37Ra, and H37Rv from three different biological samples were used for western blot analysis. (ii) Histogram depicts the relative intensity of ATP2A2 measured in case of uninfected, H37Ra, and H37Rv, respectively. Protein was normalized to the intensity of actin and quantified. (b) Diagrammatic representation of PC/PE ratio in THP-1 cells infected with either H37Ra or H37Rv. Fold change in PC/PE ratio was 6.79 in case of H37Ra while H37Rv showed only 2.76. (c) Cytosolic Ca²⁺ levels of THP-1 cells were estimated using Confocal microscopy. Fluo-4, AM (green) was used for intracellular calcium binding and measured at 494/506 nm. (d) The mean fluorescence intensity (MFI)/cell obtained for H37Ra and H37Rv infected cells was shown. The MFI for uninfected cells was . Values represent the mean ± SE of approximately 200 cells from 3 independent experiments. White bar represents H37Rv while black bar represents H37Ra.

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