Fig 1: LncRNA TNRC6C-AS1 silencing promotes the apoptosis and autophagy of TC cells through STK4/Hippo. A, The mRNA expression of Beclin1 in transfected SW579 cells normalized to GAPDH was determined by RT-qPCR. B, Western blot analysis was used to determine the protein levels of Beclin1, LC3-II/I and Casepase-3 in SW579 cells normalized to GAPDH after transfection. C, The proliferation ability of SW579 cells in each group was tested by EdU assay. D, TUNEL method was used to detect the apoptosis rate of SW579 cells. E, MDC method was used to detect the number of autophagic vesicles of SW579 cells in each group. *P < .05 vs the blank group, #P < .05 vs the shSTK4 group. The comparison among multiple groups was analysed by one-way ANOVA (mean ± standard deviation). Values were obtained from 3 independent experiments in triplicate
Fig 2: Effects of the ERß inhibitor PHTPP on autophagy-related proteins in HIBD neonatal mice pretreated with ICA. The expression of (a) Beclin1, (b) LC3, and (c) p62 in the cerebral ischemic penumbra (black box of the brain slice in the upper left corner) in neonatal HIBD mice treated with PHTPP detected by fluorescence, n = 8 mice in each group. Bar = 100 µm. (d) Representative western blot images and (e–g) quantitative analysis. ***P < 0.001, and ****P < 0.0001 compared to the HI + ICA + Vehicle group. Data are presented as the mean ± SDs, n = 6 mice in each group.
Fig 3: Effects of the autophagy inhibitor 3-MA on autophagy-related proteins in HIBD neonatal mice pretreated with ICA. Immunofluorescence detection of the expression of (a) Beclin1, (b) LC3, and (c) p62 in the ischemic penumbra (small black box in the upper left brain slice) of 3-MA-treated HIBD neonatal mice, n = 8 mice in each group. Bar = 100 µm. (d) Representative western blot images and (e–g) quantitative analysis. ***P < 0.001, and ****P < 0.0001 compared to the HI + ICA + Vehicle group. Data are presented as the mean ± SDs, n = 6 mice in each group.
Fig 4: Risperidone confers an inhibitory effect on differentiation and autophagy and a promotive effect on apoptosis of osteoblasts in vitro. A the mRNA expression of OPG, collagen I and RANKL in MC3T3-E1 cells determined by RT-qPCR. B–C Western blot analysis of differentiation-related protein (OPG, collagen I and RANKL) expression in MC3T3-E1 cells. The band intensity was assessed. D representative images of ALP staining in MC3T3-E1 cells. E, statistical results of ALP positive rate in MC3T3-E1 cells. F representative images of alizarin red S staining in MC3T3-E1 cells. G statistical results of integrated optical density (IOD) of alizarin red S in MC3T3-E1 cells. H representative images of autophagosome in MC3T3-E1 cells assessed by immunofluorescence. I Western blot analysis of autophagy-related protein (LC3 II/I, Beclin1, and p62) expression in MC3T3-E1 cells. The band intensity was assessed. J Western blot analysis of apoptosis-related protein (cleaved PARP1, cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9) expression in MC3T3-E1 cells. K apoptosis rate in MC3T3-E1 cells evaluated by flow cytometry. *p < 0.05, compared with MC3T3-E1 cells treated with normal culture medium. The cell experiments are repeated 3 times
Fig 5: mRNA and protein expression levels of LC3 and BECN1 in the natural aging group and D-gal-induced mimetic aging group. (Aa-c) Reverse transcription-quantitative polymerase chain reaction was used to determine the changes in the mRNA expression levels in the D-gal group compared with those in the age-matched NS group. (Ad-f) Western blot analysis was used to detect the protein expression levels of LC3-II and BECN1. ß-actin was included as a loading control. (B) Changes in LC3 and BECN1 protein expression with aging were demonstrated using immunofluorescence, and the nuclei were stained with DAPI. Data are expressed as mean + standard error of the mean. *P<0.05, **P<0.01 and ***P<0.001 vs. the age-matched NS subgroup; ^P<0.01 vs. the 3-month-old NS subgroup; #P<0.001 vs. the 3-month-old D-gal subgroup. Scale bar, 100 mm at magnification, ×200 and 50 mm at magnification, ×600. NS, normal saline; D-gal, D-galactose; BECN1, Beclin 1.
Supplier Page from Abcam for Anti-Beclin 1 antibody