Comparison between CD24 antibody treatment and CD24 expression manipulation.

<p>A) and B) Comparison of varying cisplatin IC-50 values in UM-SCC-10B on CD24 antibody treatment, CD24 knockdown variant as compared with the control, standard error of mean (t-test, n = 3). C) and D) Comparison of varying cisplatin IC-50 values in UM-SCC-15s on CD24 antibody treatment, CD24 knockdown variant as compared with the control, standard error of mean (t-test, n = 3). E) and F) Comparison of varying cisplatin IC-50 values in UM-SCC-74B on CD24 antibody treatment, CD24 overexpression variants as compared with the control, standard error of mean (t-test, n = 3).</p

Modulation of CD24 expression directly affects the gene expression of various pro-survival genes critical in cisplatin resistance.

<p>A) and B) Down regulation of CD24 in CD24 rich UM-SCC-10B and UM-SCC-15s leads to significant down regulation of critical pro-survival genes, standard error of mean (t-test, n = 3). C) Up regulation of CD24 in CD24 deficient UM-SCC-74B leads to significant up regulation of critical pro-survival genes, standard error of mean (t-test, n = 3). D) Graph of the number of orospheres formed by the CD24 knockdown variants of UM-SCC-10B and UM-SCC-15s before and after an IC-50 dose of cisplatin treatment, standard error of mean (t-test, n = 3). E) Graph of the number of orospheres formed by the CD24 overexpressing variants of UM-SCC-74B before and after an IC-50 dose of cisplatin treatment, standard error of mean (t-test, n = 3).</p

No Correlation between CD44% and cisplatin sensitivity.

<p>A) Hematoxylin and Eosin staining of three laryngeal carcinoma cell lines. B) Cisplatin IC-50 calculated for the three laryngeal carcinoma lines. C) Flow cytometry analysis of membrane bound CD44 on the three laryngeal carcinoma lines and D) graphical representation of CD44% with standard error of mean (t-test, n = 3). E) Graph of the number of orospheres formed with standard error of mean (t-test, n = 3).</p

CD24 plays a functional role in determining cisplatin sensitivity of UM-SCC-74B.

<p>A) CD24 expression Lentivector map purchased from GeneCopoeia (US). B) RNA level confirmation of CD24 up-regulation in UM-SCC-74B, standard error of mean (t-test, n = 3). C) Protein level confirmation of CD24 up-regulation in UM-SCC-74B. D) Cisplatin IC-50 value CD24 overexpressing UM-SCC-74B rises significantly as compared to the parental UM-SCC-74B. E) Graph showing cisplatin IC-50 value of CD24 overexpressing UM-SCC-74B rises significantly as compared to the parental UM-SCC-74B, standard error of mean (t-test, n = 3).</p

Real time PCR primers to probe gene expression in lines with modified CD24 expression levels.

<p>Real time PCR primers to probe gene expression in lines with modified CD24 expression levels.</p

CD24 expression and its correlation with cisplatin treatment outcome.

<p>A) Tumor volumes of UM-SCC-10B vector control and its CD24 knockdown clone UM-SCC-10B CD24KD treated with Saline (Control) and Cisplatin (Test), standard error of mean (t-test, n = 8). B) Tumor volumes of UM-SCC-15s vector control and its CD24 knockdown clone UM-SCC-15s CD24KD treated with Saline (Control) and Cisplatin (Test), standard error of mean (t-test, n = 8). C) Tabulation tumor volumes measured for UM-SCC-10B and UM-SCC-10BCD 24KD; graph of the final volume of the tumors and their statistical measure (standard error of mean). D) C) Tabulation tumor volumes measured for UM-SCC-15s and UM-SCC-15s CD24KD; graph of the final volume of the tumors and their statistical measure (standard error of mean).</p

Correlation between CD24% and cisplatin sensitivity.

<p>A) Tabulation showing that CD24+ is an indicator of poor outcome in multiple cancers. B) Tabulation showing that the positive or negative expression of CD24 is the discerning factor in isolating cancer stem cells/ tumor propagating cells in various cancers. C) Flow cytometry analysis of membrane bound CD24 and CD44 of the three laryngeal carcinoma lines show an increasing fraction of CD24% and D) Graphical representation of CD24% with R² = 0.804 showing a positive linear correlation with increasing cisplatin resistance in the three laryngeal carcinoma lines. E) Primary and secondary orospheres formed by the CD24+ (green) and CD24– (red) fractions. F) Graph of the number of orospheres formed as shown in E) with standard error of mean (t-test, n = 3). G) CD24+ residual fraction increase in proportion as it remains viable even after an IC-50 dose of cisplatin treatment. H) Graph with standard error of mean (t-test, n = 3) of the percent increase in proportion of the residual viable CD24+ fraction after an IC-50 dose of cisplatin treatment.</p