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Fig. 2 | BMC Molecular and Cell Biology

Fig. 2

From: PGRMC1 effects on metabolism, genomic mutation and CpG methylation imply crucial roles in animal biology and disease

Fig. 2

PGRMC1 phosphorylation status does not affect P4-dependent resistance to doxorubicin toxicity or resistance to AG-205-induced cell death. a P4 reduces cell proliferation of cells expressing all PGRMC1-HA proteins. The panel shows boxplots of viable cells for n = 6 replicates. The viability of cells pretreated with P4 (dark boxes) or DMSO vehicle control (light boxes) after 23 h were allowed to grow a further 24 h and the level of MTT formazan was quantified as a proxy for viable cell numbers. “% cells after 48 hr” is presented relative to the signal obtained after adherence for 3 h. No significant differences were observed between non-P4 treated cell pairings, except MP v. TM (p = 0.021, 2 way ANOVA). Pair wise comparisons of P4-treated MP vs any of P4-treated WT, DM or TM, or of +/−P4 treatment for WT, DM and TM, were significantly different at the p < 0.00001 level (2 way ANOVA). MP cells +/− P4 were not different (p = 0.55, two way ANOVA). Considering only P4-treated cells, MP differed in P4 response from all other cell types (p < 1 × 10–10), and WT differed marginally from TM (p = 0.034) by one way ANOVA and post-hoc Bonferroni. b P4-protection of MP cells from doxorubicin-induced cell death is facilitated by over-expression of PGRMC1-HA proteins (WT, DM & TM). Cells were grown as in (a), except at t0 doxorubicin (Dox) was added at the indicated concentrations, followed by 24 h incubation. Because of altered cell proliferation during pretreatment with P4 (a), all signals at t0 were expressed as a percentage of the averaged control sample without Dox at t0 to assess the effects of P4. Data points represent the averages ± s.d. of n = 6 replicates. c Boxplots of area under the curve (AUC) results for all data points with dox addition from (b). Two way ANOVA dox treatments were statistically significant (F = 1292.237, df = 1, df2 = 40, p < 1 = e-8), with Partial Eta Squared indicating 97% effect size in the data. Considering pairwise comparisons of cell classes, DM v. TM (p = 0.014) and WT v. DM (p = 0.05) were significantly different. Pairwise comparison between the AUC levels for +/− P4 for each cell class yielded no difference for MP, but p < 1 × 10–8 for other cell types, +/− P4. One way ANOVA post-hoc Bonferroni pairwise comparisons for -P4 cells revealed that samples within dotted boxes did not differ significantly, whereas all pairwise p values were less than those indicated for comparisons between boxes

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