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ATCC endocervical end1

Transwell assay model. Monolayers of endocervical <t>(End1),</t> ectocervical (Ect1) and Vaginal (Vk2) epithelial cells grown in 24-well inserts for 24 hours in a liquid-liquid interface to obtain functional monolayers. The functional monolayers were treated with calcitriol at 1x10 -8 M or EtOH 0.1% at the basal side for 24 hours to resemble uptake of this hormone from circulation. Transmigration assays were performed for three hours, adding 1ng/mL of free-cell R5-tropic HIV-1 (Bal) to monolayers of epithelial cells at the upper chamber (Apical side). Activated CD4 + T cells were used as targets of infection at the lower chamber (Basal side) for evaluating infectiveness of transmigrated virions (a ratio of 1.5 epithelial cells to 1 CD4 T cells was used during the 3 hours co-culture of these cells); the remaining virions, at the upper chamber (untransmigrated), were also evaluated for their infectiveness of activated CD4 + T cells. The infection cultures for transmigrated and untransmigrated virions were maintained by 84 hours and infection was determined by detection of p24 by flow cytometry.

Endocervical End1, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression"

Article Title: Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2026.1758656

Transwell assay model. Monolayers of endocervical (End1), ectocervical (Ect1) and Vaginal (Vk2) epithelial cells grown in 24-well inserts for 24 hours in a liquid-liquid interface to obtain functional monolayers. The functional monolayers were treated with calcitriol at 1x10 -8 M or EtOH 0.1% at the basal side for 24 hours to resemble uptake of this hormone from circulation. Transmigration assays were performed for three hours, adding 1ng/mL of free-cell R5-tropic HIV-1 (Bal) to monolayers of epithelial cells at the upper chamber (Apical side). Activated CD4 + T cells were used as targets of infection at the lower chamber (Basal side) for evaluating infectiveness of transmigrated virions (a ratio of 1.5 epithelial cells to 1 CD4 T cells was used during the 3 hours co-culture of these cells); the remaining virions, at the upper chamber (untransmigrated), were also evaluated for their infectiveness of activated CD4 + T cells. The infection cultures for transmigrated and untransmigrated virions were maintained by 84 hours and infection was determined by detection of p24 by flow cytometry.

Figure Legend Snippet: Transwell assay model. Monolayers of endocervical (End1), ectocervical (Ect1) and Vaginal (Vk2) epithelial cells grown in 24-well inserts for 24 hours in a liquid-liquid interface to obtain functional monolayers. The functional monolayers were treated with calcitriol at 1x10 -8 M or EtOH 0.1% at the basal side for 24 hours to resemble uptake of this hormone from circulation. Transmigration assays were performed for three hours, adding 1ng/mL of free-cell R5-tropic HIV-1 (Bal) to monolayers of epithelial cells at the upper chamber (Apical side). Activated CD4 + T cells were used as targets of infection at the lower chamber (Basal side) for evaluating infectiveness of transmigrated virions (a ratio of 1.5 epithelial cells to 1 CD4 T cells was used during the 3 hours co-culture of these cells); the remaining virions, at the upper chamber (untransmigrated), were also evaluated for their infectiveness of activated CD4 + T cells. The infection cultures for transmigrated and untransmigrated virions were maintained by 84 hours and infection was determined by detection of p24 by flow cytometry.

Techniques Used: Transwell Assay, Functional Assay, Transmigration Assay, Infection, Co-Culture Assay, Flow Cytometry

Comparison of the percentage of HIV-1 infection of CD4 + T cells with virions from the apical or basal sides of End1, Ect1 and Vk2 epithelial monolayers in the absence of treatment. Comparison of the frequency of infected p24 + CD4 + T cells with virions from the basal side (lower chamber) and apical side (upper chamber) of transwell inserts containing untreated End1 (basal, light blue dots; apical, dark blue dots), Ect1 (basal, light green dots; apical, dark green dots) or Vk2 monolayers (basal, light pink dots; apical, dark pink dots). Infection of CD4 + T cells in absence of epithelial cells (No-EpCs) included as a positive control of viral infectious capacity and cell susceptibility to infection (gray dots). Comparison between infectiveness of HIV-1 virions from the apical and basal sides of each epithelial monolayer were made using the Ratio paired t-test, and comparisons among the three cell types for the HIV-1 infection with virions from the apical or basal compartments were done using One-way ANOVA. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments).

Figure Legend Snippet: Comparison of the percentage of HIV-1 infection of CD4 + T cells with virions from the apical or basal sides of End1, Ect1 and Vk2 epithelial monolayers in the absence of treatment. Comparison of the frequency of infected p24 + CD4 + T cells with virions from the basal side (lower chamber) and apical side (upper chamber) of transwell inserts containing untreated End1 (basal, light blue dots; apical, dark blue dots), Ect1 (basal, light green dots; apical, dark green dots) or Vk2 monolayers (basal, light pink dots; apical, dark pink dots). Infection of CD4 + T cells in absence of epithelial cells (No-EpCs) included as a positive control of viral infectious capacity and cell susceptibility to infection (gray dots). Comparison between infectiveness of HIV-1 virions from the apical and basal sides of each epithelial monolayer were made using the Ratio paired t-test, and comparisons among the three cell types for the HIV-1 infection with virions from the apical or basal compartments were done using One-way ANOVA. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments).

Techniques Used: Comparison, Infection, Positive Control

Frequency of HIV-1-infected CD4 + T cells and MFI of p24 in CD4 + T cells following the culture with virions retained at the apical side (upper chamber) of the VitD- or EtOH-conditioned genital epithelial monolayers. Comparison of the percentage of p24 + CD4 + T cells following contact with untransmigrated HIV-1 Bal (1ng of p24) exposed to VitD- or EtOH-treated End1 (A) , Ect1 (C) or Vk2 monolayers (E) . Comparison of the MFI of expression of p24 in CD4 + T cells following contact of virions with calcitriol- or EtOH-treated End1 (B) , Ect1 (D) or Vk2 monolayers (F) . All the epithelial cells were treated with VitD or EtOH for 24 hours before co-cultures. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments). The percentage of HIV-1 reduction by VitD compared to EtOH treatment is depicted in each figure. Comparison between treatments were made using the Ratio paired t-test, *p≤ 0.05; **p≤ 0.01; ***p≤ 0.001.

Figure Legend Snippet: Frequency of HIV-1-infected CD4 + T cells and MFI of p24 in CD4 + T cells following the culture with virions retained at the apical side (upper chamber) of the VitD- or EtOH-conditioned genital epithelial monolayers. Comparison of the percentage of p24 + CD4 + T cells following contact with untransmigrated HIV-1 Bal (1ng of p24) exposed to VitD- or EtOH-treated End1 (A) , Ect1 (C) or Vk2 monolayers (E) . Comparison of the MFI of expression of p24 in CD4 + T cells following contact of virions with calcitriol- or EtOH-treated End1 (B) , Ect1 (D) or Vk2 monolayers (F) . All the epithelial cells were treated with VitD or EtOH for 24 hours before co-cultures. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments). The percentage of HIV-1 reduction by VitD compared to EtOH treatment is depicted in each figure. Comparison between treatments were made using the Ratio paired t-test, *p≤ 0.05; **p≤ 0.01; ***p≤ 0.001.

Techniques Used: Infection, Comparison, Expressing

Frequency of HIV-1-infected CD4 + T cells and MFI of p24 in CD4 + T cells following the culture with virions transmigrated through the VitD- or EtOH-conditioned epithelial monolayers (lower chamber). Comparison of the percentage of p24 + CD4 + T cells following culture with HIV-1 Bal (1ng of p24) transmigrated through VitD- or EtOH-treated End1 (A) , Ect1 (C) or Vk2 monolayers (E) . Comparison of the MFI of expression of p24 in CD4 + T cells following the transmigration of virions through VitD- or EtOH-treated End1 (B) , Ect1 (D) or Vk2 monolayers (F) . Comparison of the frequency of p24 + CD4 + T cells infected with virions from the basal side (lower chamber) and apical side (upper chamber) of VitD-treated End1(light blue dots represent the basal compartment; dark blue dots represent the apical compartment), Ect1 (light green dots, basal; and dark green dots, apical) or Vk2 (light pink dots, basal; and dark pink dots, apical) monolayers (G) . All the epithelial cells were treated with VitD or EtOH for 24 hours before co-cultures. A ratio of 1.5 epithelial cells to 1 CD4 T cells was used during the 3 hours co-culture of these cells. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments). The percentage of HIV-1 reduction by VitD compared to EtOH treatment is depicted in each figure. Comparison between VitD and EtOH treatments and comparison between infectious virions at the apical and basal sides of each epithelial monolayer were made using the Ratio paired t-test, *p≤ 0.05; **p≤ 0.01; ***p≤ 0.001; ****p<0.0001.

Figure Legend Snippet: Frequency of HIV-1-infected CD4 + T cells and MFI of p24 in CD4 + T cells following the culture with virions transmigrated through the VitD- or EtOH-conditioned epithelial monolayers (lower chamber). Comparison of the percentage of p24 + CD4 + T cells following culture with HIV-1 Bal (1ng of p24) transmigrated through VitD- or EtOH-treated End1 (A) , Ect1 (C) or Vk2 monolayers (E) . Comparison of the MFI of expression of p24 in CD4 + T cells following the transmigration of virions through VitD- or EtOH-treated End1 (B) , Ect1 (D) or Vk2 monolayers (F) . Comparison of the frequency of p24 + CD4 + T cells infected with virions from the basal side (lower chamber) and apical side (upper chamber) of VitD-treated End1(light blue dots represent the basal compartment; dark blue dots represent the apical compartment), Ect1 (light green dots, basal; and dark green dots, apical) or Vk2 (light pink dots, basal; and dark pink dots, apical) monolayers (G) . All the epithelial cells were treated with VitD or EtOH for 24 hours before co-cultures. A ratio of 1.5 epithelial cells to 1 CD4 T cells was used during the 3 hours co-culture of these cells. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments). The percentage of HIV-1 reduction by VitD compared to EtOH treatment is depicted in each figure. Comparison between VitD and EtOH treatments and comparison between infectious virions at the apical and basal sides of each epithelial monolayer were made using the Ratio paired t-test, *p≤ 0.05; **p≤ 0.01; ***p≤ 0.001; ****p<0.0001.

Techniques Used: Infection, Comparison, Expressing, Transmigration Assay, Co-Culture Assay

Transcriptional expression of VitD pathway genes in genital epithelial cells after VitD treatment. Fold changes of expression of CYP24A1 at 6h (A) and 24 h (B) of VitD or EtOH treatments of End1, Ect1 and Vk2 cells, and comparison between 6h and 24h time points (C) . Fold changes of expression of VDR at 6h (D) and 24 h (E) of VitD treatment of End1, Ect1 and Vk2, and comparison between 6h and 24h time points (F) . Fold-change in transcript (2 -ΔΔCt (effects) ) was calculated using ΔΔC t(effects) = ΔC t(target gene of treatment group) - ΔC t (target gene of untreated group) . Average threshold cycle (C t ) of 3 technical PCR replicates of the target gene was standardized against the average C t of the 18S rRNA (internal input reference) of the corresponding sample, termed ΔC t (target gene) . Shown was the data of 4 independent experiments (i.e., 4 transwell filters per treatment group, per cell line). Ratio paired t test was used. *p≤ 0.05; **p≤ 0.01.

Figure Legend Snippet: Transcriptional expression of VitD pathway genes in genital epithelial cells after VitD treatment. Fold changes of expression of CYP24A1 at 6h (A) and 24 h (B) of VitD or EtOH treatments of End1, Ect1 and Vk2 cells, and comparison between 6h and 24h time points (C) . Fold changes of expression of VDR at 6h (D) and 24 h (E) of VitD treatment of End1, Ect1 and Vk2, and comparison between 6h and 24h time points (F) . Fold-change in transcript (2 -ΔΔCt (effects) ) was calculated using ΔΔC t(effects) = ΔC t(target gene of treatment group) - ΔC t (target gene of untreated group) . Average threshold cycle (C t ) of 3 technical PCR replicates of the target gene was standardized against the average C t of the 18S rRNA (internal input reference) of the corresponding sample, termed ΔC t (target gene) . Shown was the data of 4 independent experiments (i.e., 4 transwell filters per treatment group, per cell line). Ratio paired t test was used. *p≤ 0.05; **p≤ 0.01.

Techniques Used: Expressing, Comparison

Expression of cathelicidin (CAMP) gene and protein in genital epithelial cells following VitD treatment. Fold changes of the CAMP RNA transcripts at 6h (A) and 24 h (B) following VitD- or EtOH- treatment of End1, Ect1 and Vk2, and comparison between 6h and 24h time points (C) . Fold-change in transcript (2 -ΔΔCt (effects) ) was calculated using ΔΔC t(effects) = ΔC t(target gene of treatment group) - ΔC t (target gene of untreated group) , as described in <xref ref-type=

Figure 6 . Shown was the data of 4 independent experiments (i.e., 4 transwell filters per treatment group, per cell line). Ratio paired t tests were used. *p≤ 0.05; **p≤ 0.01. The amount of cathelicidin protein released into the apical side of End1, Ect1 and Vk2 monolayers was assessed using ELISA (D) . Three independent experiments (3 transwell inserts) for each cell type were performed. The supernatant from the apical side of the monolayers were pooled and concentrated using speed vacuum; concentrations shown were adjusted according to concentration factor, to nanogram of cathelicidin per ml of supernatant. " title="... h (B) following VitD- or EtOH- treatment of End1, Ect1 and Vk2, and comparison between 6h and ..." property="contentUrl" width="100%" height="100%"/>
Figure Legend Snippet: Expression of cathelicidin (CAMP) gene and protein in genital epithelial cells following VitD treatment. Fold changes of the CAMP RNA transcripts at 6h (A) and 24 h (B) following VitD- or EtOH- treatment of End1, Ect1 and Vk2, and comparison between 6h and 24h time points (C) . Fold-change in transcript (2 -ΔΔCt (effects) ) was calculated using ΔΔC t(effects) = ΔC t(target gene of treatment group) - ΔC t (target gene of untreated group) , as described in Figure 6 . Shown was the data of 4 independent experiments (i.e., 4 transwell filters per treatment group, per cell line). Ratio paired t tests were used. *p≤ 0.05; **p≤ 0.01. The amount of cathelicidin protein released into the apical side of End1, Ect1 and Vk2 monolayers was assessed using ELISA (D) . Three independent experiments (3 transwell inserts) for each cell type were performed. The supernatant from the apical side of the monolayers were pooled and concentrated using speed vacuum; concentrations shown were adjusted according to concentration factor, to nanogram of cathelicidin per ml of supernatant.

Techniques Used: Expressing, Comparison, Enzyme-linked Immunosorbent Assay, Concentration Assay

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Journal: Frontiers in Immunology

Article Title: Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression

doi: 10.3389/fimmu.2026.1758656

Figure Lengend Snippet: Transwell assay model. Monolayers of endocervical (End1), ectocervical (Ect1) and Vaginal (Vk2) epithelial cells grown in 24-well inserts for 24 hours in a liquid-liquid interface to obtain functional monolayers. The functional monolayers were treated with calcitriol at 1x10 -8 M or EtOH 0.1% at the basal side for 24 hours to resemble uptake of this hormone from circulation. Transmigration assays were performed for three hours, adding 1ng/mL of free-cell R5-tropic HIV-1 (Bal) to monolayers of epithelial cells at the upper chamber (Apical side). Activated CD4 + T cells were used as targets of infection at the lower chamber (Basal side) for evaluating infectiveness of transmigrated virions (a ratio of 1.5 epithelial cells to 1 CD4 T cells was used during the 3 hours co-culture of these cells); the remaining virions, at the upper chamber (untransmigrated), were also evaluated for their infectiveness of activated CD4 + T cells. The infection cultures for transmigrated and untransmigrated virions were maintained by 84 hours and infection was determined by detection of p24 by flow cytometry.

Article Snippet: Immortalized endocervical (End1) (ATCC CRL-2615 TM ), ectocervical (Ect1) (ATCC CRL-2614 TM ) and Vaginal (Vk2) (ATCC CRL-2616 TM ) epithelial cells were used in this study.

Techniques: Transwell Assay, Functional Assay, Transmigration Assay, Infection, Co-Culture Assay, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression

doi: 10.3389/fimmu.2026.1758656

Figure Lengend Snippet: Comparison of the percentage of HIV-1 infection of CD4 + T cells with virions from the apical or basal sides of End1, Ect1 and Vk2 epithelial monolayers in the absence of treatment. Comparison of the frequency of infected p24 + CD4 + T cells with virions from the basal side (lower chamber) and apical side (upper chamber) of transwell inserts containing untreated End1 (basal, light blue dots; apical, dark blue dots), Ect1 (basal, light green dots; apical, dark green dots) or Vk2 monolayers (basal, light pink dots; apical, dark pink dots). Infection of CD4 + T cells in absence of epithelial cells (No-EpCs) included as a positive control of viral infectious capacity and cell susceptibility to infection (gray dots). Comparison between infectiveness of HIV-1 virions from the apical and basal sides of each epithelial monolayer were made using the Ratio paired t-test, and comparisons among the three cell types for the HIV-1 infection with virions from the apical or basal compartments were done using One-way ANOVA. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments).

Article Snippet: Immortalized endocervical (End1) (ATCC CRL-2615 TM ), ectocervical (Ect1) (ATCC CRL-2614 TM ) and Vaginal (Vk2) (ATCC CRL-2616 TM ) epithelial cells were used in this study.

Techniques: Comparison, Infection, Positive Control

Journal: Frontiers in Immunology

Article Title: Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression

doi: 10.3389/fimmu.2026.1758656

Figure Lengend Snippet: Frequency of HIV-1-infected CD4 + T cells and MFI of p24 in CD4 + T cells following the culture with virions retained at the apical side (upper chamber) of the VitD- or EtOH-conditioned genital epithelial monolayers. Comparison of the percentage of p24 + CD4 + T cells following contact with untransmigrated HIV-1 Bal (1ng of p24) exposed to VitD- or EtOH-treated End1 (A) , Ect1 (C) or Vk2 monolayers (E) . Comparison of the MFI of expression of p24 in CD4 + T cells following contact of virions with calcitriol- or EtOH-treated End1 (B) , Ect1 (D) or Vk2 monolayers (F) . All the epithelial cells were treated with VitD or EtOH for 24 hours before co-cultures. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments). The percentage of HIV-1 reduction by VitD compared to EtOH treatment is depicted in each figure. Comparison between treatments were made using the Ratio paired t-test, *p≤ 0.05; **p≤ 0.01; ***p≤ 0.001.

Article Snippet: Immortalized endocervical (End1) (ATCC CRL-2615 TM ), ectocervical (Ect1) (ATCC CRL-2614 TM ) and Vaginal (Vk2) (ATCC CRL-2616 TM ) epithelial cells were used in this study.

Techniques: Infection, Comparison, Expressing

Journal: Frontiers in Immunology

Article Title: Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression

doi: 10.3389/fimmu.2026.1758656

Figure Lengend Snippet: Frequency of HIV-1-infected CD4 + T cells and MFI of p24 in CD4 + T cells following the culture with virions transmigrated through the VitD- or EtOH-conditioned epithelial monolayers (lower chamber). Comparison of the percentage of p24 + CD4 + T cells following culture with HIV-1 Bal (1ng of p24) transmigrated through VitD- or EtOH-treated End1 (A) , Ect1 (C) or Vk2 monolayers (E) . Comparison of the MFI of expression of p24 in CD4 + T cells following the transmigration of virions through VitD- or EtOH-treated End1 (B) , Ect1 (D) or Vk2 monolayers (F) . Comparison of the frequency of p24 + CD4 + T cells infected with virions from the basal side (lower chamber) and apical side (upper chamber) of VitD-treated End1(light blue dots represent the basal compartment; dark blue dots represent the apical compartment), Ect1 (light green dots, basal; and dark green dots, apical) or Vk2 (light pink dots, basal; and dark pink dots, apical) monolayers (G) . All the epithelial cells were treated with VitD or EtOH for 24 hours before co-cultures. A ratio of 1.5 epithelial cells to 1 CD4 T cells was used during the 3 hours co-culture of these cells. Each paired points represent one blood donor for the source of CD4 T cells, for a total of six biological replicates (i.e., 6 independent experiments). The percentage of HIV-1 reduction by VitD compared to EtOH treatment is depicted in each figure. Comparison between VitD and EtOH treatments and comparison between infectious virions at the apical and basal sides of each epithelial monolayer were made using the Ratio paired t-test, *p≤ 0.05; **p≤ 0.01; ***p≤ 0.001; ****p<0.0001.

Article Snippet: Immortalized endocervical (End1) (ATCC CRL-2615 TM ), ectocervical (Ect1) (ATCC CRL-2614 TM ) and Vaginal (Vk2) (ATCC CRL-2616 TM ) epithelial cells were used in this study.

Techniques: Infection, Comparison, Expressing, Transmigration Assay, Co-Culture Assay

Journal: Frontiers in Immunology

Article Title: Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression

doi: 10.3389/fimmu.2026.1758656

Figure Lengend Snippet: Transcriptional expression of VitD pathway genes in genital epithelial cells after VitD treatment. Fold changes of expression of CYP24A1 at 6h (A) and 24 h (B) of VitD or EtOH treatments of End1, Ect1 and Vk2 cells, and comparison between 6h and 24h time points (C) . Fold changes of expression of VDR at 6h (D) and 24 h (E) of VitD treatment of End1, Ect1 and Vk2, and comparison between 6h and 24h time points (F) . Fold-change in transcript (2 -ΔΔCt (effects) ) was calculated using ΔΔC t(effects) = ΔC t(target gene of treatment group) - ΔC t (target gene of untreated group) . Average threshold cycle (C t ) of 3 technical PCR replicates of the target gene was standardized against the average C t of the 18S rRNA (internal input reference) of the corresponding sample, termed ΔC t (target gene) . Shown was the data of 4 independent experiments (i.e., 4 transwell filters per treatment group, per cell line). Ratio paired t test was used. *p≤ 0.05; **p≤ 0.01.

Article Snippet: Immortalized endocervical (End1) (ATCC CRL-2615 TM ), ectocervical (Ect1) (ATCC CRL-2614 TM ) and Vaginal (Vk2) (ATCC CRL-2616 TM ) epithelial cells were used in this study.

Techniques: Expressing, Comparison

Journal: Frontiers in Immunology

Article Title: Vitamin D boosts HIV-1 resistance in female genital epithelial cells by enhancing antiviral cathelicidin expression

doi: 10.3389/fimmu.2026.1758656

Figure Lengend Snippet: Expression of cathelicidin (CAMP) gene and protein in genital epithelial cells following VitD treatment. Fold changes of the CAMP RNA transcripts at 6h (A) and 24 h (B) following VitD- or EtOH- treatment of End1, Ect1 and Vk2, and comparison between 6h and 24h time points (C) . Fold-change in transcript (2 -ΔΔCt (effects) ) was calculated using ΔΔC t(effects) = ΔC t(target gene of treatment group) - ΔC t (target gene of untreated group) , as described in Figure 6 . Shown was the data of 4 independent experiments (i.e., 4 transwell filters per treatment group, per cell line). Ratio paired t tests were used. *p≤ 0.05; **p≤ 0.01. The amount of cathelicidin protein released into the apical side of End1, Ect1 and Vk2 monolayers was assessed using ELISA (D) . Three independent experiments (3 transwell inserts) for each cell type were performed. The supernatant from the apical side of the monolayers were pooled and concentrated using speed vacuum; concentrations shown were adjusted according to concentration factor, to nanogram of cathelicidin per ml of supernatant.

Article Snippet: Immortalized endocervical (End1) (ATCC CRL-2615 TM ), ectocervical (Ect1) (ATCC CRL-2614 TM ) and Vaginal (Vk2) (ATCC CRL-2616 TM ) epithelial cells were used in this study.

Techniques: Expressing, Comparison, Enzyme-linked Immunosorbent Assay, Concentration Assay

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