Fig 2: Estradiol contributes to autophagy in chondrocytes by mediating the downregulation of ASIC1a. (A and B) The expression of ASIC1a decreased in a time- and concentration-dependent manner following stimulation with 0–2000 nmol/mL estradiol for 0 h to 48 h in chondrocytes. (C and D) The expression of Beclin1, LC3 and p62 protein in chondrocytes in a time- and concentration-dependent manner following stimulation with 0–2000 nmol/mL estradiol for 0 h to 48 h. (E) Estradiol induced expression levels of ASIC1a in chondrocytes cultured in pH 6.0 medium for 24 h with or without E2 (500 nmol/mL) were analyzed by immunofluorescence. Representative views from each group are presented (original magnification, ×400). Data are presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 versus 0 nmol/mL group; #P < 0.05, ##P < 0.01 versus 0 h group.

Fig 3: Effect of P. falciparum infection during pregnancy in placental autophagy-related genes expression.(A-C) Placental mRNA expression of the selected autophagy-related genes ULK1 (A), BECN1 (B), and MAP1LC3B (C) in non-infected (NI) and P. falciparum-infected (Pf-INF) women. Results represent qPCR estimates relative to NI and normalized by GAPDH, endogenous control. Data sets on placental mRNA levels from P. falciparum-infected women were afterwards stratified according to placental malaria (PM) status as placental malaria negative (Pf-INF PM-) or positive (Pf-INF PM+) and plotted for ULK1 (D), BECN1 (E), and MAP1LC3B (F). (G) Heatmap matrixes of NI and Pf-INF placentas pairwise Spearman correlation coefficients (rs) between mRNA levels and histologic and immunologic parameters. Data are represented as whiskers boxplots where the bottom and the top of the box are the first and third quartiles, the line inside the box is the median and the whiskers represent the minimum and the maximum values (A-F), and heatmaps containing rs in a colour range from dark blue (rs = -1) to dark red (rs = 1) with statistically significant correlations enhanced as bold values (G). The differences between groups were determined by the Mann-Whitney U test (A-C), Kruskal-Wallis test with Dunn’s post-test for multiple comparisons (D-F), and Spearman’s rank-order non-parametric test for correlations (G). P values < 0.05 were considered as representing statistically significant differences. BW–birth weight, PW–placental weight, HZ–hemozoin, SNA–syncytial nuclear aggregates, NECR–necrosis, VASC–vascularity, LEU–leukocytes, MO–monocytes.

Fig 4: Overexpression of BCL2L10 inhibited autophagy of Hep3B cells by binding to BECN1. (A) The expression of BCL2L10 and BECN1 after transfection in four different groups was detected by qRT-PCR. (B) The cell viability of pcDNA3.1-BECN1 group was the weakest, while the cell viability of pcDNA3.1-BCL2L10 group was the strongest. (C) Overexpression of BECN1 facilitated the accumulation of LC3B puncta in Hep3B cells as detected by immunofluorescence. (D) The expression of LC3B-II/LC3B-I in pcDNA3.1-BCL2L10 group was decreased while the expression of LC3B-II/LC3B-I in pcDNA3.1-BECN1 group was increased. The changes of P62 had a contrary trend with changes of LC3B-II/LC3B-I. (E) Degradation of P62 was little observed in the presence of Bafilomycin A1 (an inhibitor of late-phase autophagy), while LC3B-II/LC3B-I expression was increased in Hep3B cells. * P <0.05.

Fig 5: Function annotations for BCL2L10 / BECN1 in HCC. (A-B) Hierarchical cluster analysis of the 10 most up and down regulated mRNAs. In the heat maps, green represents genes that are down-regulated whereas red represents genes that are up-regulated. (C) Plot of ten most enriched KEGG pathways in HCC. Pathways are ordered by normalized enrichment score (NES). Percentage beside the bar indicates the proportion of differential genes in pathway gene set. (D) STRING co-expression network for BCL2L10/ BECN1 and their related signaling pathways.