The goal of this subsection is to assess the robustness of our results to the inclusion of an additional control for both outcome variables.
When referees asked for an additional control experiment recently, I went in for the day and sorted it.
For an additional control, the cells were incubated together with free human IgG F ab′ 2.
The discrepancy argues for an additional control mechanism, such as mRNA localization, transcript stability, post-translational regulation, and protein degradation.
Cells within SL-biofilms were also fixed by adding 37% formaldehyde (Fisher Chemical, Waltham, MA, F79-500) to a final concentration of 4% and incubating for 1 h prior to imaging as an additional control for cellular vitality.
Immunostaining using an antibody to cytokeratin 18 (DAKO) was used as an additional control for the staining method.
In addition a glmM PCR (Table 1) was carried out [ 16], both as an additional control for H. pylori identification and quality check of the isolated DNA (positive PCR control).
As an additional control for the potentially confounding effects of scratching, we used a system in which a cell-free gap was produced by a removable spacer without mechanical effects on cells or coating material.
As an additional control for "self priming" during PCR [22], DENV2 isolate 62663 first was treated with 10 µg/ml of RNAse A for 10 min at 37°C to eliminate any unpackaged RNA or other RNA contaminant.
As an additional control for specificity, PD-L2 was also stained after MIH5 or MAC219 treatments.
In addition, the SMARTpool ON-TARGETplus CACNA1C siRNA (Dharmacon, Cat. No. L-006123-00-0005) was used an additionalnal control for all experiments where SMARTpool siRNA were used.
