Journal: Neural Regeneration Research

Article Title: PTEN inhibitor bisperoxovanadium protects against noise-induced hearing loss

doi: 10.4103/1673-5374.358606

Figure Lengend Snippet: bpV prevents noise-induced reductions in PI3K-Akt signaling in OHCs. (A, B) Representative images showing the reduced levels of PI3K-p85α and -p110α (red, Alexa Fluor 594) in OHCs after noise exposure, as detected by immunofluorescence staining. Pretreatment with bpV protected against these effects. Scale bars: 20 μm. (C, D) Quantification of PI3K-p85α and -p110α levels in OHCs in noise (PTS) and noise + bpV (PTS + bpV) groups. (E, F) Representative images and quantification analyses showed that p-Akt levels were reduced in OHCs after noise exposure. (G) mRNA expression levels of PI3K and Akt were detected by real-time PCR in control, noise, and noise + bpV groups. (H) Protein expression of PI3K-p85α, -p110α, p-Akt and Akt were determined by western blotting. (I–K) Quantification of PI3K-p85α and -p110α relative to GAPDH and of p-Akt/Akt. Expression of p110α decreased after noise exposure compared with that in the control group. There was no change in total Akt level between the three groups. Data are presented as the mean ± SD. The experiment was repeated three times. * P < 0.05, ** P < 0.01, *** P < 0.001 (unpaired Student’s t -test). bpV: Bisperoxovanadium; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IHC: inner hair cell; OHC: outer hair cell; p-Akt: phospho-Akt; PCR: polymerase chain reaction; PI3K: phosphoinositide-3 kinase; PTS: permanent threshold shift.

Article Snippet: The primary antibodies were monoclonal rabbit anti-PTEN (Cell Signaling Technology, Cat# 9188, RRID: AB_2253290), monoclonal rabbit anti-PI3K-p110α (Cell Signaling Technology, Cat# 4249, RRID: AB_2165248), polyclonal rabbit anti-PI3K-p85α (Cell Signaling Technology, Cat# 4292, RRID: AB_329869), monoclonal rabbit anti-phospho-Akt (Ser 473) (Cell Signaling Technology, Cat# 4060, RRID: AB_2315049) and polyclonal rabbit anti-4 hydroxynonenal (4HNE; Abcam, Cambridge, UK, Cat# ab46545, RRID: AB_722490).

Techniques: Immunofluorescence, Staining, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Argon preconditioning protects neuronal cells with a Toll-like receptor-mediated effect

doi: 10.4103/1673-5374.355978

Figure Lengend Snippet: Argon preconditioning reduces rotenone-induced protein expression of Bax and increases Bcl-2, but the effect is abolished by OxPAPC. (A, C) Densitometric analysis of western blots for Bax (A) and Bcl-2 (C) protein expression. Between-group comparisons were performed with a one-way analysis of variance using the post hoc Holm Sidak test ( n = 6; mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001). (B, D) Representative western blot images of six independent experiments that showed similar results demonstrating the influence of rotenone, argon preconditioning, and inhibition with OxPAPC on Bax (B) and Bcl-2 (D).

Article Snippet: After protein transfer to polyvinylidene difluoride membranes (Immobilon-P; Millipore, Schwalbach, Germany), the membranes were blocked with 5% skim milk in Tween20/PBS (TBST) or BSA (bovine serum albumin) and incubated in the recommended dilution of protein-specific antibody (p44/42 MAP kinase (phospho-ERK1/2 [extracellular-signal regulated kinase]) (Thr202/Tyr204), rabbit, 1:2000, Cell Signaling Technology, Danvers, MA, USA, Cat# 9101, RRID: AB_331646; phospho-NF-κB (Ser536) (93H1), rabbit, 1:1000, Cell Signaling Technology, Cat# 3033, RRID: AB_331284; phospho-Akt (protein kinase B) antibody (Ser473), rabbit, 1:1000, Cell Signaling Technology, Cat# 9271, RRID: AB_329825; Bax antibody, rabbit, 1:1000, Cell Signaling Technology, Cat# 2772, RRID: AB_10695870; Bcl-2 antibody, rabbit, 1:1000, Cell Signaling Technology, Cat# 2876, RRID: AB_2064177; and cleaved caspase-3 antibody (Asp175), rabbit, 1:500, Cell Signaling Technology, Cat# 9661, RRID: AB_2341188; NF-E2-related factor 2 (Nrf2) antibody [nuclear factor erythroid 2-related factor 2, phosphor-S40], rabbit, 1:60000, Abcam, Cat# ab76026, RRID: AB_1524049).

Techniques: Expressing, Western Blot, Inhibition