rabbit anti simkania negevensis antibodies  (Absolute Biotech Inc)

Journal: Applied and Environmental Microbiology

Article Title: Temperature Affects the Host Range of Rhabdochlamydia porcellionis

doi: 10.1128/aem.00309-23

Figure Lengend Snippet: Permissivity of arthropod and mammalian cell lines to R. porcellionis. (A) The y axis represents the fold change of the number of genome copies per microliter relative to the initial time point. The results are shown as the mean and standard deviation from three biological replicates. (B) R. porcellionis in mammalian and arthropod cell lines at 6 days postinfection. Growth could be observed only in Sf9 cells. The reticulate bodies do not appear to be grouped in an inclusion and seem to be replicating directly in the cytoplasm. The enlarged bodies in the C6/36 cell line are likely aberrant bodies. Bacteria appear to have been internalized in all the other cell lines but failed to replicate. White arrows indicate enlarged bacteria in C6/36 cells and internalized EBs in the other cell lines. Cells were stained with concanavalin A (red), DAPI (blue), and anti- Simkania antibody (green); bar, 10 μm.

Article Snippet: The coverslips were then incubated at room temperature for 2 h in blocking solution with rabbit anti- Simkania negevensis antibodies ( ) (dilution at 1:1,000), rabbit anti- Waddlia chondrophila antibodies ( ) (dilution at 1:1,000), or goat antibodies targeting the major outer membrane protein of Chlamydia trachomatis (dilution at 1:1,000) (LSBio, Seattle, WA, USA).

Techniques: Standard Deviation, Staining

Journal: Applied and Environmental Microbiology

Article Title: Temperature Affects the Host Range of Rhabdochlamydia porcellionis

doi: 10.1128/aem.00309-23

Figure Lengend Snippet: The growth of R. porcellionis in mammalian cells depends on temperature. (A) Growth kinetics of R. porcellionis in Sf9 cells incubated at 20, 28, 33, and 37°C. (B) Growth kinetics of R. porcellionis in mammalian cells incubated at 28°C. In both panel A and panel B, the y axis represents the fold change relative to the initial time point. (C) Doubling time of R. porcellionis in the different cell lines. Doubling times were estimated by dividing 48 h by the log 2 of the highest fold change observed between two consecutive time points. Despite the marked difference between the doubling time in Sf9 and the other cell lines, the Kruskal-Wallis test was not statistically significant ( P = 0.06). (D) IFU count of R. porcellionis grown in different cell lines at 28°C. The cells were fixed at 6 days postinfection. The tendency of infected Sf9 cells to detach from the glass coverslips could induce an underestimation of the IFU count. A one-way ANOVA revealed that there was a statistically significant difference between at least two cell lines ( P value = 0.0002). The plot shows the results of the Tukey honestly significant difference test for the pairwise comparison of the IFU count in the different cell lines (**, <0.01; ***, <0.001). (E) McCoy, A549, and Ishikawa cells infected with R. porcellionis , incubated at 28°C, and fixed at 6 days postinfection. The two enlarged bodies in McCoy and A549 cells are likely aberrant bodies (white arrows). Cells were stained with concanavalin A (red), DAPI (blue), and anti- Simkania antibodies (green). Bar, 10 μm. The results show the mean and standard deviation from three biological replicates.

Article Snippet: The coverslips were then incubated at room temperature for 2 h in blocking solution with rabbit anti- Simkania negevensis antibodies ( ) (dilution at 1:1,000), rabbit anti- Waddlia chondrophila antibodies ( ) (dilution at 1:1,000), or goat antibodies targeting the major outer membrane protein of Chlamydia trachomatis (dilution at 1:1,000) (LSBio, Seattle, WA, USA).

Techniques: Incubation, Infection, Staining, Standard Deviation

Journal: bioRxiv

Article Title: Temperature affects the host range of Rhabdochlamydia porcellionis and the infectivity of Waddlia chondrophila and Chlamydia trachomatis elementary bodies

doi: 10.1101/2022.09.02.506449

Figure Lengend Snippet: (A) Permissivity of arthropod and mammalian cell lines to R. porcellionis . The y-axis represents the log2 fold change of the number of genome copies per μL compared to the initial timepoint (mean +/− standard deviation, n=2) (B) Infectious progeny production. The supernatant from Sf9 cells infected with R. porcellionis was collected every other day (time points indicated on the x axis) and used to infect fresh Sf9 cells. The cells were then collected at zero (dots) and 6 days (triangles) post infection to quantify bacterial growth by qPCR (mean +/− standard deviation, n=2) (C) R. porcellionis in mammalian and arthropod cell lines at 6 days post-infection. Growth could only be seen in Sf9 cells. The reticulate bodies seem to replicate directly in the cytoplasm, without being enclosed in an inclusion. The enlarged bodies in the C6/36 cell lines are likely aberrant bodies. Bacteria appear to have been internalized in all the other cell lines, but failed to replicate. White arrows indicate enlarged bacteria in C6/36 cells and internalized EBs in the other cell lines. Cells were stained with concanavalin A (red), DAPI (blue) and anti- Simkania antibody (green), scale bar: 10 μm.

Article Snippet: The coverslips were then incubated at room temperature for 2 hours in blocking solution with rabbit anti-Simkania negevensis antibodies ( ) (dilution at 1:1000), rabbit anti- Waddlia chondrophila antibodies ( ) (dilution 1:1000) or goat antibodies targeting the major outer membrane protein of Chlamydia trachomatis (dilution 1:1000) (LSBio, Seattle, USA).

Techniques: Standard Deviation, Infection, Staining

Journal: bioRxiv

Article Title: Temperature affects the host range of Rhabdochlamydia porcellionis and the infectivity of Waddlia chondrophila and Chlamydia trachomatis elementary bodies

doi: 10.1101/2022.09.02.506449

Figure Lengend Snippet: R. porcellionis grows in mammalian cells incubated at 28 °C (A) Growth kinetics of R. porcellionis in Sf9 incubated at 20, 28, 33 and 37 °C. (B) Growth kinetics of R. porcellionis in mammalian cells, incubated at 28 °C. In both (A) and (B) , the y-axis represent the log2 fold change compared to time 0 (mean +/− standard deviation, n=3) (C) IFU count in different cell lines infected with the same suspension of R. porcellionis (mean +/− standard deviation, n=3). All cells were all grown at 28 °C. A one-way ANOVA revealed that there was a statistically significant difference between at least two cell lines (p-value=0.0002). The plot also shows the results of Tukey HSD test for the pairwise comparison of the different cell lines (** < 0.01, *** < 0.001). (D) McCoy, A549 and Ishikawa cells infected with R. porcellionis and incubated at 28 °C. The two enlarged bodies in McCoy and A549 cells are likely aberrant bodies (white arrow). Cells were fixed 6 days post infection and stained with concanavalin A (red), DAPI (blue) and anti- Simkania antibodies (green). Scale bar: 10 μm.

Article Snippet: The coverslips were then incubated at room temperature for 2 hours in blocking solution with rabbit anti-Simkania negevensis antibodies ( ) (dilution at 1:1000), rabbit anti- Waddlia chondrophila antibodies ( ) (dilution 1:1000) or goat antibodies targeting the major outer membrane protein of Chlamydia trachomatis (dilution 1:1000) (LSBio, Seattle, USA).

Techniques: Incubation, Standard Deviation, Infection, Staining