For indirect immunofluorescence of HeLa cells for endogenous proteins, cells were grown on coverslips for 16 h, followed by processing for immunofluorescence as described above.
Lower back skin samples were fixed with 4% paraformaldehyde for either 30 min on ice followed by processing for frozen sectioning, or for 4 h at room temperature followed by processing for paraffin sectioning.
Further, the pyro-synthesis adopts a continuous sample processing technique in contradiction to the challenging batch-to-batch processing followed for commercial solid-state or sol-gel reactions to obtain LiFePO4.
For ionizing radiation treatment flies in standard food vials were irradiated with 5666Rads/hour for 16 hours using a Cesium source (Grammacell 40-Cesium 137, Atomic Energy, Canada) at the USC Norris Cancer Center facility, and then transferred to 1% sucrose vials for two days followed by processing for RNA.
Cells were cultured to ∼70% confluence and medium was replaced with fresh medium containing either vehicle (0.1% DMSO) or 10 μM Red-Br-nos for the noted times, followed by processing for immunofluorescence microcopy, flow cytometry or immunoblotting as described previously.
For comparison, the commercially available controls of CaCO3, Ca3(PO4)2, and HAP described earlier were diluted and washed in either double-distilled water (shown in Fig. 13J, K, L, respectively) or DMEM (data not shown), followed by processing for FTIR analysis.
In order to examine mitochondrial protein solubility in situ, we lightly fixed cells prior to extraction in low concentrations of non-ionic detergent, followed by processing for electron microscopy.
Controls of CaCO3, Ca3(PO4)2, and HAP were diluted and washed in either double-distilled water (shown in Fig. 14J, K, L, respectively) or DMEM (data not shown), followed by processing for micro-Raman analysis.
To determine whether other rRNA processing steps were affected by DDX21 reduction, we followed rRNA processing for 50 minutes.
At the end of each treatment time, cells were collected after a brief incubation with trypsin EDTA followed by processing for cell cycle analysis as reported earlier [ 14].
