Figure S11. Additional characterization of L. monocytogenes infections of stable caveolin-1 and epsin-1 knockdown cell lines.
(A) Scatter plot of the raw data values (area of infection [µm2]) from Figure 4F. The average area of infection values are as follows: 262164µm2 [Control shRNA] and 76823µm2 [Cav-1 shRNA]. ***, P < 0.0001 (unpaired parametric two-tailed t tests [with Welch’s correction]).
(B) Quantification of internalized L. monocytogenes following 8 hour infection foci experiments using HeLa cells stably expressing non-targeting control shRNA or caveolin-1-targeting shRNA [Cav-1 shRNA]. Bacterial load counts from 3 separate experiments (where each experiment was also run in triplicate) were obtained and plotted as a bar graph of the percent internalized bacteria (relative to control, [± s.d.]). The percentage of internalized bacteria (relative to control, ± s.d.) are 100 % (Control shRNA) and 101 % (Cav-1 shRNA).
(C) The average abundance of membrane protrusions, presented as a bar graph (normalized relative to control, ± s.e.m.) of the ratio of total enumerated protrusions divided by total enumerated bacterially-generated actin structures (actin clouds, comet/rocket tails and membrane protrusions). In total, 30 cells per genotype (from 3 independent experiments) were analyzed. The average amount of protrusions generated are 100 % [Control shRNA] and 96% [Cav-1 shRNA] cells.
(D) Quantification of the length of CD147-positive membrane invaginations generated in HeLa cells stably expressing non-targeting control shRNA [Control shRNA] or caveolin-1-targeting shRNA [Cav-1 shRNA]. Results are presented as a bar graph of the membrane invagination length (± s.e.m.]). In total, 30 and 28 membrane invaginations were measured from Control shRNA and Cav-1 shRNA cells respectively (15 cells per genotype; 3 independent experiments). The average invagination lengths were 8.33 µm and 4.43 µm. ***, P < 0.0001 (unpaired parametric two-tailed t tests [with Welch’s correction]).
(E) Scatter plot of the raw data values (area of infection [µm2]) from Figure 5E. The average area of infection values are as follows: 124585µm2 [Control siRNA] and 94439µm2 [Epsin-1 siRNA]. ***, P < 0.0001 (unpaired Mann Whitney U test).
(F) Percent values (relative to control) of infection foci area from Figure 5E but plotted with standard error of the mean [s.e.m.]. ***, P < 0.0001 (unpaired parametric two-tailed t tests [with Welch’s correction]).
