Fig. 4

Drug screening with CD10 as a marker identified dexamethasone and retinoic acid, which exhibit stimulatory and inhibitory effects on adipogenesis, respectively. a Dexamethasone (10 μM) was added under three conditions such as (i) D-2 to D12 (pre-induction and adipogenic stimulation), (ii) D0 to D12 (only adipogenic stimulation), and (iii) no addition. The presence and absence of dexamethasone are shown as solid and dotted lines, respectively. b Representative merged images of AdipoRed staining of lipids (in green) and Hoechst 33342 staining of nuclei (in blue) are shown in CD10 KD, CD10 OE, S40, and S8 ASCs. The scale bar represents 100 μm. c The LDs are quantified as mentioned earlier as a read-out of adipogenesis levels. Statistical analysis was performed using Student’s paired t test (n = 3) between conditions (i) and (ii), ** p < 0.01, *p < 0.05, non-significant (ns) represents p value > 0.05. d Relative mRNA levels of CD10 are quantified under the indicated conditions of dexamethasone addition normalized to RPL27. Fold expression changes are compared to the subject S40 (ii). Pre-treatment with dexamethasone significantly increased CD10 expression and subsequently lipid accumulation except for the CD10 KD line. Statistical analysis was performed using Student’s paired t test (n = 3) between condition (i) and (ii), **p < 0.01, *p < 0.05, non-significant (ns) represents p value > 0.05. Each value is the mean ± SEM from three independent replicates. e Representative merged images of AdipoRed staining of lipids (in green) and Hoechst 33342 staining of nuclei (in blue) are compared between CD10 OE and control ASCs to see the effect of retinoic acid (RA) on adipogenesis. Equivalent amount of either DMSO or RA (10 μM) was added. The scale bar represents 100 μm. f LDs was quantified based on MATLAB programming. Addition of RA significantly reduced LDs accumulation. g Relative mRNA levels of CD10 in presence or absence of RA were quantified. Fold expression changes are compared to C (OE) values. RA significantly reduced CD10 expression, and CD10 OE does not rescue RA-mediated adipogenic impairment. Each value is the mean ± SEM from three independent replicates. Significance was assessed by using Student’s paired t test: ***p < 0.001