Fig 1.
Phase-contrast images of buffalo MECs culture.
A) Mixed population of epithelial and fibroblast cells, B) Formation of islands by purified MECs, at low density seeding, C) Cobble stone morphology shown by confluent MECs, D) Post confluent stage of MECs forming dome structures (arrow), E) MECs from passage 5 forming papillate structures, F) Floating dead cells due to occurrence of contact inhibition during post confluency stage.
Fig 2.
Immunocytostaining for expression of cytoskeletal markers in buffalo MECs A) Fluorescent image of Cytokeratin 18 showing intermediate filaments running in bundles with interconnections between cells, B) Light image of Cytokeratin 18, C) Fluorescent image of buffalo MECs stained for Vimentin showing filament degradation, D) Light image of Vimentin.
Fig 3.
Cell viability pattern in heat stressed buffalo MECs using trypan blue dye exclusion method.
CTR-unstressed (control) MECs.
Fig 4.
Evaluation of cellular proliferation in unstressed (control) and heat stressed buffalo MECs using MTT assay.
CTR-unstressed (control); TRT- heat stress treated MECs.
Fig 5.
Proportions of early apoptotic, apoptotic and dead cells in unstressed (CTR) and heat stressed treated (TRT) buffalo MECs during recovery period after heat stress.
Fig 6.
Evaluation of cellular apoptosis based on flow cytometric analysis in unstressed (CTR) and heat stressed (HS) buffalo MECs using Annexin FITC/7-AAD dyes.
For each graph, the quadrants display distribution of viable (bottom left), early apoptotic (bottom right), apoptotic (upper right) and dead (upper left) cells.
Fig 7.
Bar graph showing differentially expressed genes after heat stress in buffalo MECs at different fold change (2, 3, 5 and 10).
Fig 8.
Bar graph showing up- and down-regulated genes at each time point (Fold change 3) in heat stressed buffalo MECs.
Fig 9.
Venn diagrams showing the distribution of genes identified as heat stress responsive at 3 fold change, and the overlapping genes identified as most commonly expressed at all-time points of heat stress treatment in buffalo MECs, (A) most commonly up-regulated (153 genes), (B) most commonly down-regulated (8 genes) at all-time points post heat stress.
Fig 10.
Venn diagram showing number of DEG at 30 min, 2 h, 4 h, 8 h, 12 h, 16 h, and 24 h with respect to unstressed (CTR) at fold change > = 3.0.
Fig 11.
Venn diagram showing number of genes induced at 30 min, 2 h, 4 h, 8 h, 12 h, 16 h, and 24 h with respect to unstressed (CTR) at fold change > = 3.0.
Table 1.
List of top 50 genes up-regulated in heat stressed buffalo MECs (Fold change > = 3.0).
Table 2.
List of top 50 genes down-regulated in heat stressed buffalo MECs (Fold change > = 3.0).
Table 3.
List of genes classified in major functional categories during post heat stress (relative to control) in buffalo MECs.
Table 4.
List of significant GO terms obtained from REVIGO analysis.
Fig 12.
GO categories for biological process enriched across commonly up-regulated 153 genes (relative to control).
Fig 13.
Cytokines & Inflammatory response pathway; shows the significant affected genes (yellow color) across all time points post heat stress.