Fig. 4

Expression levels of classic nPRs and non-classic mPRs/PAQRs are modulated by mPR-specific PRG actions and the CSC in nPR(+) T47D cells. A RNA expression levels of classic nPRs and mPRs (PAQR5-9) are influenced by mPR-specific PRG actions. After mPR-specific PRG treatment (20 µM each) for 48 h, RNA expression levels of classic nPRs, major mPRs (PAQR5-9) and PGRMC1 were measured (n = 3). B Protein expression levels of mPRs (PAQRs) are suppressed by mPR-specific PRG actions (20 µM each) for 48 h (n = 3). C Protein expression levels of mPRs (PAQRs) are mainly suppressed by silencing CCM1 and CCM3 genes. After silencing all three CCMs for 48 h, protein expression levels of mPRs (PAQR5-9) were performed (n = 3). D RNA expression levels of major mPRs (PAQRs, PGRMC1) are mainly modulated by CCM2. After silencing all three CCMs for 48 h, altered RNA expression of major mPRs were measured (triplicates per experiment, n = 3). E RNA expression levels of nPRs is significantly increased by silencing mPR (PAQRs) genes. After silencing mPR (PAQR7-9) genes for 48 h, RNA expression levels of nPRs were observed (triplicate per experiment, n = 3). F Decreased protein expression of PAQR7 by silencing nPRs under mPR-specific PRG actions. nPRs deficient T47D cells compared to scramble controls were stained utilizing IHC approaches with HRP/DAB (n = 4). G Both CCM1 and CCM3 proteins are stabilized to mPR-specific PRG actions after silencing mPRs in T47D cells. T47D cells were treated to silence major mPRs (PAQR5/7/8) for 24 h, followed by mPR-specific PRG treatment (20 µM each) for an additional 48 h (n = 4). Relative RNA expression changes were measured by qPCR (Fold changes) and normalized to scramble control (red line, triplicates per experiment, n = 3). Relative expression levels of proteins were measured through quantification of band intensities and normalized against either α-actinin (ACTN1) or β-actin (ACTB) followed by SC controls (red line). In all bar plots, red line is the control baseline for fold change measurements (−/+). **, *** above bar indicates P ≤ 0.01 or 0.001 for paired t-test, respectively