Additional file 3: Figure S2. of Multiplexed, targeted profiling of single-cell proteomes and transcriptomes in a single reaction

Standard protein probe curves using lysed and diluted MCF7 cells. Two-fold dilutions of population lysate were backloaded into the C1 IFC and processed according to the PEA/STA protocol. Shown here are the PEA measurements with the y-axis values representing ∆Ct, which are calculated as the signal over a lysis buffer only control. Certain assays (e.g. EIF4EBP1_P) display a “hook” effect. The effect occurs when the concentration of a target protein exceeds a threshold beyond which PEA probes begin to occupy separate target molecules as opposed to the same one. This results in a reduction of signal due to a reduction in the number of proximal events. Each data point plotted is the average of eight separate capture sites in the C1 IFC with error bars showing the standard error of the mean. Gray (green) dashes show the level above which the probability for a detection event being real is p = 0.01 (0.05). Points used for fitting the red trend line are colored blue, the background plot color indicates which treatment cells were taken from (0 hr = purple, 24 hr = green, 48 hr = blue). Probes are categorized as (a) usable or (b) unusable. (PDF 557 kb)