Figure 4.

Modulation of intracellular calcium and mitochondrial activity by H2S and the effect of NaHS mediated Calcium influx on CXCR4 expression. (a) Graphs depict the MFI of Fluo-3AM indicative of intracellular calcium measured in LSK cells. The experiment was done in PBS with or without CaCl₂ in the presence or absence of NaHS and Nifedipine (NF, L-type calcium channel blocker). (b) intracellular calcium levels before and after NaHS/NF treatment in LSK population when the cells were incubated in the IMDM media (containing pre-added CaCl₂, phenol red free) (c) the effect of H₂S on intracellular calcium levels in HEK cell line in the presence and absence of CaCl₂ indicating the effect of H2S not only in LSK fraction of mice but in the other cells also. (d) Fold change in CXCR4 expression in BMMNCs in vehicle/NaHS/BAPTA-AM treated cells demonstrating the role of calcium in NaHS mediated increase in CXCR4 expression. (e) Line graphs indicating the gradual rise in MFI of Rh123, a mitochondrial membrane potential dye after 6,12 and 24 h in NaHS/vehicle treated cells (LSK fraction). (f) mitochondrial mass in LSK proportion using Mitotraker green (g) Mitochondrial superoxide level in vehicle/NaHS treated LSK population using Mitosox red indicator at 3 different time points. (h) summary of the proposed mechanism. Data are expressed as fold change in MFI (wrt control) for n = 3 and all the statistical comparisons were done using One-way ANOVA and Bonferroni posttest