Fig. 2. Pharmacological inhibition of autophagy with chloroquine (CQ) impairs steroidogenesis in luteinized granulosa cells and corpus luteum tissue samples.
A Representative blots for indicated proteins after treatment of the luteinized granulosa cells with hCG w/wo chloroquine (CQ) at indicated concentrations. Densitometric quantification is indicated to the right of the blots. Mean ± SD, N = 6 biological replicates analyzed using one-way ANOVA, with Tukey’s test for multiple comparisons. B Representative confocal images of the cells stained for LC3 (green) and lysotracker (red) 24 h after treatment with hCG w/wo CQ at indicated concentrations. Quantification of the signal intensities and co-localizations of the signals are indicated to the right of the images. Nuclei stained with DAPI. Scale bars represent 20 μm. Mean ± SD, N = 6 biological replicates analyzed using one-way ANOVA, with Tukey’s test for multiple comparisons. C Representative graphic bar indicates progesterone (P4) production of the cells before and 24 h after treatment with hCG w/wo CQ at indicated concentrations. Mean ± SD, N = 6 biological replicates analyzed using one-way ANOVA, with Tukey’s test for multiple comparisons. D Histological section of corpus luteum (CL) tissue sample after hematoxylin-eosin staining. Scale bar represents 50 μm. E Representative blots for indicated proteins 24 h after treatment of the CL tissue samples with hCG w/wo CQ at indicated concentrations. Densitometric quantification is indicated to the right of the blots. Mean ± SD, N = 5 biological replicates analyzed using one-way ANOVA, with Tukey’s test for multiple comparisons. F Representative graphic bar indicates progesterone (P4) production of the CL tissue samples before and 24 h after treatment with hCG (10 IU/ml) w/wo CQ (60 μM). Mean ± SD, N = 5 biological replicates analyzed using one-way ANOVA, with Tukey’s test for multiple comparisons.