Figure 3.
Fip200−/−B cells showed decreased proliferation, plasma differentiation, and survival in vitro. B cells isolated from WT or B-Fip200−/− mouse spleens were cultured with IL4 (10 ng/ml) + LPS (5 μg/ml). (A and B) Expression levels of p62 (A) and LC3 (B) were detected in WT and Fip200−/− B cells upon stimulation at 0, 4, 16, 20, and 24 h. Representative data from one of at least two experimental replicates are shown. (C) Representative plots and the corresponding quantifications of plasma cells of WT and Fip200−/− B cells after 3 days of culture. Data are combined from two independent experiments with at least three mice in each group, and samples are run in triplicate. Significant (α = 0.05) P values were determined by unpaired, two-tailed Student’s t test; ****P < 0.0001. (D) WT and Fip200−/− B cells expressing Blimp-GFP were stimulated by IL4+LPS. Blimp-GFP+ (day 2) or Blimp-GFP+CD138+ (day 3) populations were checked by FACS. N = 5 biological replicates, with representative data shown from one mouse. (E and F) Representative plots (E) and the corresponding quantifications (F) of the IRF4hiPAX5lo population of WT and Fip200−/− B cells on days 1–3 of IL4 + LPS activation. Two independent experiments were performed. Representative data from one experiment with cells originating from four mice per group with two cultures generated per mouse are shown, except for KO, day 3, from which only one culture was obtained. Significant P values were determined by an unpaired t test. Left to right: ***P = 0.0002, ***P = 0.0002, **P = 0.0040. (G) Proliferation of WT and Fip200−/− B cells upon stimulation by IL4+LPS was detected by FACS at days 1–3. Left to right: ***P = 0.0003, ***P = 0.0002. (H) Representative plots and the corresponding quantifications of IRF4hiPAX5lo population in proliferated populations (CTVlo) of WT or Fip200−/− B cells on day 2 of IL4 + LPS activation. Two independent experiments were performed. Representative data from one experiment with cells originating from four mice per group with two cultures generated per mouse are shown. Significant P values were determined by an unpaired t test. ***P = 0.0002. (I) WT and Fip200−/− B cells were stimulated by IL4+LPS, and cell survival rate was detected by FACS on days 1–3. Left to right: **P = 0.0096, ***P = 0.0001, ****P < 0.0001 (unpaired t test). N = 3 biological replicates, with representative data shown from one mouse. (J) Cleaved caspase-3 was detected in WT and Fip200−/− B cells after stimulation by IL4+LPS at days 2–3. N = 2 biological replicates, with representative data shown from one mouse. Source data are available for this figure: SourceData F3.

Fip200 −/− B cells showed decreased proliferation, plasma differentiation, and survival in vitro. B cells isolated from WT or B-Fip200−/− mouse spleens were cultured with IL4 (10 ng/ml) + LPS (5 μg/ml). (A and B) Expression levels of p62 (A) and LC3 (B) were detected in WT and Fip200−/− B cells upon stimulation at 0, 4, 16, 20, and 24 h. Representative data from one of at least two experimental replicates are shown. (C) Representative plots and the corresponding quantifications of plasma cells of WT and Fip200−/− B cells after 3 days of culture. Data are combined from two independent experiments with at least three mice in each group, and samples are run in triplicate. Significant (α = 0.05) P values were determined by unpaired, two-tailed Student’s t test; ****P < 0.0001. (D) WT and Fip200−/− B cells expressing Blimp-GFP were stimulated by IL4+LPS. Blimp-GFP+ (day 2) or Blimp-GFP+CD138+ (day 3) populations were checked by FACS. N = 5 biological replicates, with representative data shown from one mouse. (E and F) Representative plots (E) and the corresponding quantifications (F) of the IRF4hiPAX5lo population of WT and Fip200−/− B cells on days 1–3 of IL4 + LPS activation. Two independent experiments were performed. Representative data from one experiment with cells originating from four mice per group with two cultures generated per mouse are shown, except for KO, day 3, from which only one culture was obtained. Significant P values were determined by an unpaired t test. Left to right: ***P = 0.0002, ***P = 0.0002, **P = 0.0040. (G) Proliferation of WT and Fip200−/− B cells upon stimulation by IL4+LPS was detected by FACS at days 1–3. Left to right: ***P = 0.0003, ***P = 0.0002. (H) Representative plots and the corresponding quantifications of IRF4hiPAX5lo population in proliferated populations (CTVlo) of WT or Fip200−/− B cells on day 2 of IL4 + LPS activation. Two independent experiments were performed. Representative data from one experiment with cells originating from four mice per group with two cultures generated per mouse are shown. Significant P values were determined by an unpaired t test. ***P = 0.0002. (I) WT and Fip200−/− B cells were stimulated by IL4+LPS, and cell survival rate was detected by FACS on days 1–3. Left to right: **P = 0.0096, ***P = 0.0001, ****P < 0.0001 (unpaired t test). N = 3 biological replicates, with representative data shown from one mouse. (J) Cleaved caspase-3 was detected in WT and Fip200−/− B cells after stimulation by IL4+LPS at days 2–3. N = 2 biological replicates, with representative data shown from one mouse. Source data are available for this figure: SourceData F3.

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