Fig 2: Role of atorvastatin in attenuating radiotherapy- (RT-) caused injury and apoptosis of intestinal epithelial cells. (a) Representative hematoxylin-eosin (H&E) staining images of jejunum tissue sections from four groups. Scale bar = 100 μm. (b) Modified Chiu's injury score is presented for each group. Data are presented as mean ± standard error (SE) (n = 8). ∗∗P < 0.01. (c) Immunohistochemistry (IHC) analysis was used to assess the expression of cleaved caspase-3 and cleaved PARP in jejunum tissue sections. Scale bar = 100 μm. (d) The protein expression levels of cleaved caspase-3 and cleaved PARP were analyzed by western blot analysis. The bar graph represents the relative ratios of the cleaved protein to the noncleaved protein. The data are presented as mean ± SE of three independent experiments. The relative ratio of the control group is arbitrarily presented as 1. ∗P < 0.05.

Fig 3: The ROCK1 activation inhibitor Y-27632 improves the symptoms of MPTP-induced PD mice by inhibiting Drp1-dependent aberrant mitochondrial fission and apoptosis.a The substantia nigra pars compacta (SNpc) of the midbrain and the striatum were prepared and used to detect the expression of ROCK1 and CF of ROCK1 using western blot analysis. b The latency for the mice to fall from the rotarod was recorded. c SNpc and striatal tissues from each group were fixed, dehydrated, and subjected to tyrosine hydroxylase (TH, a marker of dopaminergic nerve cells) staining for immunohistochemical analysis. Scale bars: 200 μm. d The number of TH-positive dopaminergic nerve cells in three randomly selected slices from each mouse was measured by Adobe Photoshop CC. e The expression of TH in the SNpc and striatum was determined by western blot analysis. f Immunohistochemical staining for C-Cas 3 and C-PARP in the SNpc and striatum are shown. Scale bars: 20 μm. g The expression of C-Cas 3 and C-PARP was determined by western blot analysis. h SNpc and striatal tissues from each group were subjected to p-Drp1 (Ser 600) staining for immunohistochemical analysis. Scale bars: 20 μm. i The expression of p-Drp1 (Ser 600) was also determined by western blot analysis. j The expression of Drp1 in mitochondrial lysates (M) was determined by western blot analysis. The data are expressed as the means ± S.D. (n = 3). ***P < 0.001

Fig 4: Early lesions developed in AKT/TAZ mice display an inflammatory infiltrate. (A) Two weeks after hydrodynamic injection of AKT and TAZ, numerous transfected cells (indicated by black arrows) were encircled by an inflammatory infiltrate. The transfected cells and the inflammatory cells could be better appreciated at higher magnification (B), with the inflammatory cells characterized by small spherical nuclei and forming a fence around the transfected cells. The transfected cells displayed elevated TAZ (C) and CK19 (D) immunoreactivity. The inflammatory cells consisted of lymphocytes, characterized by immunoreactivity for CD4 and CD45 markers (E, F), and macrophages (immunoreactivity for the F4/80 marker) (G). Some small cells encircled by the inflammatory reaction showed signs of apoptosis, as revealed by positive immunoreactivity of apoptotic bodies (indicated by black arrows) for cleaved Caspase 3 (H) and cleaved PARP (I), implying their destruction by the inflammatory response. Original magnification: 100x in (A), 400x in (B-I); scale bar: 200 μm in (A), 50 μm in (B-I).

Fig 5: The ROCK1 activation inhibitor Y-27632 attenuates MPP+-induced Drp1-dependent aberrant mitochondrial fission and apoptosis through the inhibition of Drp1 dephosphorylation/activation.a PC12 cells were treated with MPP+ (1 mM) alone or pretreated with ROCK1 activation inhibitor Y-27632 (50 µM) for 2 h and then treated with MPP+ (1 mM) for 24 h. The expression of ROCK1 and CF of ROCK1 in whole-cell lysates was determined by western blot analysis. b The expression of p-Drp1 (Ser 656) was determined by western blot analysis. c The expression of Drp1 in mitochondrial lysates (Mito) and in cytosolic fractions (Cyto) was determined by western blot analysis. d Cells were transfected with a DsRed-Mito plasmid, and the mitochondria were viewed by confocal microscopy. Scale bars: 5 μm. e Mitochondrial length was quantified using the Imaris software. f The expression of C-Cas 3 and C-PARP in whole-cell lysates was determined by western blot analysis. g, h The apoptosis rate was measured by flow cytometry using annexin V-FITC/PI staining. The data are expressed as the means ± S.D. (n = 3). ***P < 0.001