Research Article
Progressive Rotavirus Infection Downregulates Redox-Sensitive Transcription Factor Nrf2 and Nrf2-Driven Transcription Units
Figure 6
Attenuation of Nrf2/HO-1 during RV infection is independent of Nrf2 negative regulator Keap1. (a) Keap1 protein levels were checked in MA104 cells at indicated time points post RV-SA11 infection. Relative fold change of Keap1 is represented; “ns” and “” represent comparisons with respect to the mock-infected control. (b) Scrambled siRNA/Keap1 siRNA transfected MA104 cells (kept for 36 hours) were either mock infected or infected with RV-SA11 for 9 hours before cellular extract preparation followed by SDS-PAGE/immunoblot analyses for assessing protein levels of Nrf2, HO-1, and Keap1. Relative fold changes of proteins are represented; “” and “#” represent comparisons with respect to scrambled siRNA transfected mock-infected and Keap1 siRNA transfected mock-infected groups, respectively. (c) Lysates from mock-infected and RV-SA11-infected (9 hpi) MA104 cells cotreated with tBHQ (25 μM and 50 μM; added at 1 hpi) or vehicle control (ethanol) were subjected to SDS-PAGE/immunoblotting for analyzing protein levels of Nrf2 and HO-1. Relative fold changes of proteins are represented; “,” “#,” and “$” represent comparisons with respect to vehicle-treated mock-infected, tBHQ- (50 μM) treated mock-infected, and tBHQ- (25 μM) treated mock-infected groups, respectively. (d) Mock-infected and RV-SA11-infected MA104 cells were treated with tBHQ (50 μM) or vehicle control (ethanol) at the time of final media addition. Cells fixed at 9 hpi were processed for confocal microscopy to visualize Nrf2. Scale bar, 20 μM. (e) Nrf2 CTCF from each panel of (d) was shown. “” and “#” represent comparisons with respect to vehicle-treated mock-infected and tBHQ- (50 μM) treated mock-infected groups. (f) Quotients of nuclear hollowing (NHQ) of Nrf2 in tBHQ-treated+mock-infected and tBHQ-treated+RV-SA11-infected cells were represented with respect to the vehicle- (ethanol) treated+mock-infected control.
| (a) |
| (b) |
| (c) |
| (d) |
| (e) |
| (f) |