Journal: Stem Cell Research & Therapy

Article Title: Aldo-keto reductase family 1 member C1 regulates the osteogenic differentiation of human ASCs by targeting the progesterone receptor

doi: 10.1186/s13287-021-02425-3

Figure Lengend Snippet: Knockdown of AKR1C1 enhanced the osteogenic capacity and impaired the adipogenic capacity of hASCs in vitro. a Transfection efficiency showed by fluorescence microscopy. b Knockdown efficiency of AKR1C1 determined by qRT-PCR and western blot. c Knockdown of AKR1C1 enhanced the ALP activity after 7 days of osteogenic induction as shown by ALP staining and quantification. d Knockdown of AKR1C1 accelerated mineralization after 21 days of osteogenic induction as shown by ARS staining and quantification. e Knockdown of AKR1C1 promoted the mRNA expression of RUNX2 on the 7th day of osteogenic induction and BGLAP on the 14th day of osteogenic induction as determined by qRT-PCR. f The protein expression of RUNX2 tested by western blot was consistent with the result of qRT-PCR. g Knockdown of AKR1C1 inhibited lipid droplet formation after 21 days of adipogenic induction as shown by oil red O staining. h Knockdown of AKR1C1 inhibited the mRNA expression of PPARγ and CEBPα on the 7th day of adipogenic induction as determined by qRT-PCR. i The protein expression of PPARγ tested by western blot was consistent with the result of qRT-PCR. ** p < 0.01, *** p < 0.001

Article Snippet: An alkaline phosphatase (ALP) Staining Kit (CWBIO, Beijing, China) was then used to stain the cells.

Techniques: In Vitro, Transfection, Fluorescence, Microscopy, Quantitative RT-PCR, Western Blot, Activity Assay, Staining, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Aldo-keto reductase family 1 member C1 regulates the osteogenic differentiation of human ASCs by targeting the progesterone receptor

doi: 10.1186/s13287-021-02425-3

Figure Lengend Snippet: The regulation of AKR1C1 on hASCs differentiation in vitro depended on its enzyme activity. a ALP staining indicated that overexpression of AKR1C1 by wild type plasmid (WT) in the AKR1C1 knockdown cells (sh AKR1C1 -1) downregulated the ALP activity whereas mutant plasmids (E127D, H222I and R304L) had no significant influence. The result of ALP quantification was consistent with the result of ALP staining. b The results of ARS staining and quantification were consistent with the results of ALP staining and quantification. c Overexpression of AKR1C1 by wild type plasmid (WT) in the AKR1C1 knockdown cells (sh AKR1C1 -1) downregulated the mRNA expression of RUNX2 and BGLAP whereas mutant plasmids (E127D, H222I and R304L) made no significant difference. d Western blot showed that transfection of wild type plasmid and mutant plasmids upregulated the protein expression of AKR1C1. The protein expression of RUNX2 was inhibited by transfection of wildtype plasmid but not mutant plasmids. e Oil red O staining revealed that in the AKR1C1 knockdown cells (sh AKR1C1 -1), more lipid droplet formed in WT group, whereas mutant plasmids (E127D, H222I and R304L) made no significant difference. f Overexpression of AKR1C1 by wild type plasmid (WT) in the AKR1C1 knockdown cells (sh AKR1C1 -1) upregulated the mRNA expression of PPARγ and CEBPα whereas mutant plasmids (E127D, H222I, and R304L) had no significant influence. g Western blot showed that transfection of wild type plasmid and mutant plasmids upregulated the protein expression of AKR1C1. The protein expression of PPARγ was promoted by transfection of wild type plasmid but not mutant plasmids. ** p < 0.01, *** p < 0.001

Article Snippet: An alkaline phosphatase (ALP) Staining Kit (CWBIO, Beijing, China) was then used to stain the cells.

Techniques: In Vitro, Activity Assay, Staining, Over Expression, Plasmid Preparation, Mutagenesis, Expressing, Western Blot, Transfection

Journal: Stem Cell Research & Therapy

Article Title: Aldo-keto reductase family 1 member C1 regulates the osteogenic differentiation of human ASCs by targeting the progesterone receptor

doi: 10.1186/s13287-021-02425-3

Figure Lengend Snippet: Knockdown of AKR1C1 promoted osteogenesis of hASCs in vitro through targeting PGR. a , b AKR1C1 knockdown inhibited expression of PGR but not AR validated by qRT-PCR and western blot. c , d QRT-PCR and western blot indicated that overexpression of AKR1C1 by wild type plasmid (WT) in the AKR1C1 knockdown cells upregulated expression of PGR whereas mutant plasmids (E127D, H222I, and R304L) had no significant influence. e ARS staining and quantification showed PGR knockdown in ASCs accelerated mineral accumulation after 21 days of osteogenic induction. f , g QRT-PCR and western blot showed that PGR knockdown in ASCs promoted expression of RUNX2 and BGLAP after 14 days of osteogenic induction. h Expression of PGR in AKR1C1 knockdown cells slightly downregulated the ALP activity. i Expression of PGR in AKR1C1 knockdown cells slowed down mineral accumulation. j , k QRT-PCR and western blot showed that expression of PGR in AKR1C1 knockdown cells downregulated the expression of osteogenic markers (RUNX2, 7 day and BGLAP, 14 day). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: An alkaline phosphatase (ALP) Staining Kit (CWBIO, Beijing, China) was then used to stain the cells.

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Western Blot, Over Expression, Plasmid Preparation, Mutagenesis, Staining, Activity Assay